Accelerating Results - Microarray Analysis
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Transcript of Accelerating Results - Microarray Analysis
USB Product Catalog
PCRGenomics
Custom Manufacturing
responsive
ISO certified
diagnostics
life science research
Accelerating ResultsReagents. Enzymes. Biochemicals.
proven
© 2014 Affymetrix Inc. All rights reserved. FOR RESEARCH USE ONLY.
Affymetrix, Inc. USB® Products26111 Miles RoadCleveland, Ohio 44128 Tel: 888-362-2447 | 216-765-5000Fax: 800-535-0898 | 216-464-5075 [email protected] usb.affymetrix.com
Affymetrix UK Ltd. USB® ProductsVoyager, Mercury Park,Wycombe Lane, Wooburn Green,High Wycombe HP10 0HH, United KingdomTel: +44 (0)1628 55 2600Fax: +44 (0)1628 55 [email protected]
Affymetrix Pte Ltd USB® Products7 Gul Circle, #2M-01 Keppel Logistics Building Singapore 629563 Tel: +65 63957301Fax: +65 63957300 [email protected]
CATALOG ISSUE 1.14
Results.Reagents. Enzymes. Biochemicals.
usb.affymetrix.com
Life Science Research For more than 40 years, USB has been supplying labs worldwide with molecular biology reagents, enzymes, and biochemicals. We offer a wide variety of products that scientists count on to deliver reliable, repeatable results. Our products work the first time so you can confidently advance your research.
Diagnostics We meet the regulatory compliance needs of our diagnostic customers with our ISO 13485 certified site. Our processes are suited for IVD assays and kits, with validated equipment and production practices. We offer rigorous change control capabilities and comprehensive documentation. Top diagnostic companies include our products in their kits and workflows, and in their service labs.
Custom & Bulk Manufacturing From our site in Cleveland, Ohio, we can customize any USB product to your specific need. Our expert staff can assist with bulk sizes, specialized packaging, or custom formulations. Let us lead the manufacturing, labeling, and kitting processes for all of your OEM components.
Product Code Index 252-270
Nucleotides 54-61
PCR Tools 6-53
Molecular Biology Enzymes 62-95
Molecular Biology Products & Kits 96-137
Biochemicals 182-251
Purification 138-155
RNA Analysis 156-167
DNA Sequencing 168-181
Custom, Bulk, and OEM4-5
Product Name Index 271-292
USB Product Catalog
ABOUT USB PRODUCTS
Welcome to our latest catalog. We hope it continues
to be a valued resource.
We’re committed to helping academic institutions,
diagnostic, biotech, and pharmaceutical companies
worldwide excel within the life sciences industry.
Our team is creative, dedicated, and passionate about
working together to achieve success. Our attention
to quality is evident in all that we do, from product
development and operations to our exceptional
technical support and customer service.
In addition to our catalog products, we can customize
our enzymes and biochemicals to fit your specific
requirements. Whether you need a bulk size or a
custom formulation, our expert staff will work within
our rigorous quality system to ensure that your needs
are met.
For the latest information on new products, or to
download protocols, information sheets, or MSDS
documents, please visit us at usb.affymetrix.com.
2 888-362-2447 | 216-765-5000 | usb.affymetrix.com
Ordering Information/ISO Certification
Ordering Information
Orders:Phone: 1-888-362-2447 or 216-765-5000
(8:30 a.m. - 5:30 p.m., E.S.T.)
Fax: 800-535-0898 or 216-464-5075
Website: usb.affymetrix.com
E-mail: [email protected]
Mail: Affymetrix, Inc. USB Products 26111 Miles Road Cleveland, Ohio 44128
In Europe:Phone: +44(0)1628 55 2600
Fax: +44(0)1628 55 2675
Website: usb.affymetrix.com
E-mail: [email protected]
Mail: Affymetrix UK Ltd.Voyager, Mercury Park, Wycombe Lane, Wooburn Green, High Wycombe HP10 0HH, United Kingdom
To ensure that orders are processed cor-rectly and expediently, please be sure to specify the following: Customer ID number Ship-to address Purchase order or credit card number Product number and name Quantity and size End-user’s name, department, and phone
number VAT identification number (if applicable)
Prices subject to change.
Technical support:In the U.S. and direct:
Phone: 1-888-362-2447 or 216-765-5000 (8:30 a.m. - 5:30 p.m., E.S.T.)
Fax: 800-535-0898 or 216-464-5075
E-mail: [email protected]
In Europe:
Phone: +44(0)1628 55 2600
Fax: +44(0)1628 55 2675
E-mail: [email protected]
MSDS requests:On-line: www.usb.affymetrix.com E-mail: [email protected]
Standing orders: Assure timely product delivery on pre-assigned dates. These are multiple, predetermined shipments of the same product or products.
Blanket orders: For added flexibility and convenience when ordering a variety of products over a given period of time.
Bulk orders: This may include large quanti-ties of lot-specific material, custom packag-ing (size, special labeling, etc.), or other unique specifications and requirements.
Shipping information:In the U.S., we use the following shipment methods:
�� All -20°C and 4°C products ship for next-day delivery in most areas.�� All ambient products ship “ground” for 2 to 3 day delivery. If you require next-day delivery for these items, please ask your Customer Service Representative.�� In addition, hazardous products ship via next-day delivery.�� Heavier bulk items can be shipped a number of different ways that best meet your needs and budget.
PRODUCT USES:CAUTION: Products of Affymetrix which are or may be drugs, food additives or diagnostic reagents, as described in the federal food, drug and cosmetic act, are for investigational use only in laboratory research animals or testing in vitro, and are not for drug, new drug, veterinary drug, food, food additive or human use. Unless otherwise indicated, all products are distributed and sold for chemical purposes only, not for drug use or for application to or ingestion by humans or for commercial horticulture use, for pesticide use, for application to or ingestion by animals or for veterinary drug use. All products sold by Affymetrix to Buyer shall be used by qualified professionals only. The burden for safe use and handling of all products sold by Affymetrix to Buyer is entirely the responsibility of Buyer and anyone who purchases the goods from Buyer and uses them. Absence of hazardous warnings does not imply non-toxicity. Any resale, distribution and/or export of products sold by seller to buyer outside the U.S.A. must be strictly in accordance with U.S. law and United Nations regulations.
Please visit www.usb.affymetrix.com/terms for the Affymetrix terms and conditions of sale.
Terms & Conditions
3For bulk or alternate pack sizes, email us at [email protected].
Ordering Information/ISO Certification
The site responsible for manufacturing and distribut-ing the USB-branded products has maintained cer-tification to an ISO standard since 1998. Originally certified to ISO 9000, we are pleased to announce that the site is now certified to ISO 13485:2003. While sharing many similarities with ISO 9001, ISO 13485 demonstrates the site’s preparedness for manufacturing and distributing products which may
be marketed and sold as in vitro diagnostic (IVD) reagents. This is an important change as the site continues to add new capabilities and capacities for Affymetrix.
It is our belief that ISO certification demonstrates our commitment to quality through all segments of the company. Customers can be assured that the employees take quality very seriously. Quality
products and services are obtained through the understanding, participation, and commitment of all employees.
For additional information on the site’s ISO certifica-tion, contact Amy Nasr, Director of Reagent QA and Regulatory Affairs: [email protected], 1-888-362-2447 ext. 5809.
ISO Certification
Affymetrix®, USB®, ExoSAP-IT®, Change-IT™, DeliverX™, FideliTaq™, Genetic Performance Certified™, Hi-Res™, HotStart-IT®, Ligate-IT™,
MagniTaq®, OptiKinase™, Prep2Seq™, PrepEase®, RapidRun™, RubyTaq™, SuperSAP™, Tested User Friendly™, and VeriQuest® are trademarks or
registered trademarks of Affymetrix, Inc. All other trademarks are the property of their respective owners.
Trademarks
Patents & Licenses
DNA Labeling Reagent (DLR) – This product may be covered by one or more of the following patents: U.S. Patent Nos. 6,864,059; 6,965,020; and 7,423,143.
ExoSAP-IT for PCR Product Clean-Up, HT ExoSAP-IT High-Throughput PCR Product Clean-Up, SBE Clean-Up Reagent, SNP-IT Clean-Up Reagent, Sequenase PCR Product Sequencing Kit – These products are covered by patents owned by Affymetrix, including U.S. Patent Nos. 6,379,940 and 6,387,634 and cor-responding patents in Canada and Japan.
ExoSAP-IT for PCR Product Clean-Up, HT ExoSAP-IT High-Throughput PCR Product Clean-Up, SBE Clean-Up Reagent, SNP-IT Clean-Up Reagent, Sequenase PCR Product Sequencing Kit, and the PCR Product Pre-Sequencing Kit – These products or portions thereof are sold under license from GE Healthcare under U.S. Patent Nos. 5,741,676 and 5,756,285.
HotStart-IT Products – These products may be covered by U.S. Patent Nos. 7,700,281; 7,951,534; and 8,404,443, and corresponding foreign patents. These products are also sold under licensing arrangements with Agilent Technologies, Inc. The purchase price of these products includes limited, nontransferable rights under U.S. Patent Nos. 5,605,824, 5,646,019 and 5,773,257 owned by Agilent Technologies to use only this amount of the product to practice the claims in said patents solely for activities of end users within the field of life sci-ence research. Further licensing information may be obtained by contacting the Business Development Department, Agilent Technologies, Inc., 11011 N. Torrey Pines Road, La Jolla, California 92037, USA.
MagniTaq Multiplex PCR Master Mix – Patent pending.
Poly(A) Tail-Length Assay Kit – Patent pending.
PrepEase Genomic DNA Isolation Kit – This product or portions thereof is manufactured under license from Case Western Reserve University and U.S. Pat-ent Number 7,687,254.
RNA Labeling Reagent (RLR) – This product may be covered by one or more of the following patents: U.S. Patent Nos. 6,864,059; 6,965,020; 7,282,327; 7,468,243; and 7,491,818.
Taq DNA Polymerase – Use of this product is covered by one or more of the following U.S. patents and corresponding patent claims outside the U.S.: 5,789,224, 5,618,711, and 6,127,155. The purchase of this product includes a limited, non-transferable immunity from suit under the foregoing patent claims for using only this amount of product for the purchaser’s own internal research. No right under any other patent claim, no right to perform any patented method, and no right to perform commercial services of any kind, including without limitation reporting the results of purchaser’s activi-ties for a fee or other commercial consideration, is conveyed expressly, by implication, or by estoppel. This product is for research use only. Diagnostic uses under Roche patents require a separate license from Roche. Further information on purchasing licenses may be obtained by contacting the Director of Licensing, Applied Biosystems, 850 Lincoln Centre Drive, Foster City, California 94404, USA.
SYBR Green Products – Use of these products are covered by one or more of the following US patents and corresponding patent claims outside the US: 5,994,056 and 6,171,785. The purchase of these products includes a limited, non-transferable immunity from suit under the foregoing patent claims for using only this amount of product for the purchaser’s own internal research. No right under
any other patent claim and no right to perform commercial services of any kind, including without limitation reporting the results of purchaser’s activi-ties for a fee or other commercial consideration, is conveyed expressly, by implication, or by estoppel. This product is for research use only. Diagnostic uses under Roche patents require a separate license from Roche. Further information on purchasing licenses may be obtained by contacting the Direc-tor of Licensing, Applied Biosystems, 850 Lincoln Centre Drive, Foster City, California 94404, USA. This product is also provided under an agreement between Molecular Probes, Inc. and Affymetrix and the manufacture, use, sale or import of this product is subject to one or more U.S. Patents and corresponding international equivalents, owned by Molecular Probes, a Life Technologies Corporation company. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product for real-time PCR conducted by the buyer, for use in research and development, which excludes use in performing testing, analysis or screening services. The buyer cannot sell or otherwise transfer (a) this product (b) its compo-nents or (c) materials made using this product or its components to a third party, whether or not such product or its components are resold for use in research and development, and shall not use this product or its components or materials made using this product or its components for therapeutic or prophylactic purposes. For information on purchas-ing a license to this product for purposes other than set forth above, contact Molecular Probes, Inc., Business Development, 29851 Willow Creek Road, Eugene, OR 97402, USA. Tel: (541) 465-8300, Fax: (541) 335-0354.
4 888-362-2447 | 216-765-5000 | usb.affymetrix.com
We deliver custom enzymes and bulk biochemicals with your complete satisfaction as our goal. Our team combines operational flexibility and personalized service with the production resources of a large organization.
Turn to USB for Custom, Bulk, and OEM solutions to fit your specific needs.
Contact us at [email protected] to get started.
USB® has been helping diagnostic, biotech, and pharmaceutical companies succeed in
the life sciences industry for more than 40 years. We can assist you with custom reagent
manufacturing, dispensing, labeling, and kitting of your critical reagent components.
Optimize your use of key staff and limited resources by outsourcing production of your
finished goods or key components to our team.
Custom, Bulk, and OEM
5For bulk or alternate pack sizes, email us at [email protected].
EnzymesLigases, kinases, phosphatases, polymerases, and Sequenase
Convenience ReagentsPre-mixed buffers and stock solutions
PCR ReagentsExoSAP-IT® PCR Cleanup reagent for both standard and high throughput,
SSB binding protein, DNA polymerases, and exo-free klenow
NucleotidesdNTPs, PCR nucleotide mixes, ddNTPs, and NTPs
BiochemicalsAgaroses, acrylamides, amino acids, sugars, substrates, and vitamins
Product PortfolioHere’s a sampling of our more than 3,000 customizable products:
Custom, Bulk, and OEM solutions ■ Custom enzymes, formulated to your special requirements■ Bulk sizes – our enzymes and biochemicals can be packaged into large quantities ■ Special fill sizes/concentrations/formulations of over 3,000 catalog products■ OEM labeling of individual components, or complete kit packaging and labeling with your brand■ General Purpose Reagents (GPR) and IVD reagents & assays to your exacting standards■ Customer-specific quality control assays
The USB team■ Enzymology expertise■ Extensive research, development, and manufacturing experience■ Long term supply agreements and licensing options ■ Experts who will work with you on site or from our reagents center of excellence
ISO 13485 compliant processes■ Rigorous quality system suited for your diagnostic requirements■ Comprehensive documentation■ Change control capabilities ■ We welcome audits of our manufacturing facilities
Custom, Bulk, and OEM
PCR ToolsPCR Amplification
End Point PCR
Long and Accurate PCR
Hot Start PCR
Multiplex PCR
Real-Time PCR (qPCR)
Real-Time Reverse Transcription
PCR (qRT-PCR)
Reverse Transcription PCR (RT-PCR)
PCR Purification
Passive Reference Dyes
7For bulk or alternate pack sizes, email us at [email protected].
PCR Tools | Table of Contents
PCR Product ReviewClear Choices for PCR 8PCR Product Selection Guide 9Instrument Compatibility with VeriQuest qPCR
Master Mixes 10
End Point PCRTaq DNA Polymerase 11Taq PCR Master Mix (2X) 12Taq PCR Kit 12RubyTaq™ DNA Polymerase 13RubyTaq PCR Master Mix (2X) 14
Long and Accurate PCRFideliTaq™ DNA Polymerase 15FideliTaq PCR Master Mix (2X) 16HotStart-IT® FideliTaq DNA Polymerase 17HotStart-IT FideliTaq PCR Master Mix (2X) 18
Hot Start PCRHotStart-IT Taq DNA Polymerase 19HotStart-IT Taq PCR Master Mix (2X) 20HotStart-IT Binding Protein 21
Multiplex PCRMagniTaq® Multiplex PCR Master Mix (2X) 22Promoter Methylation PCR Kit 24HotStart-IT FideliTaq DNA Polymerase 25
Real-Time qPCRVeriQuest® Taq DNA Polymerase 26VeriQuest SYBR® Green qPCR Master Mix (2X) 27VeriQuest SYBR Green qPCR Master Mix with
Fluorescein (2X) 29VeriQuest Fast SYBR Green qPCR Master
Mix (2X) 30VeriQuest Fast SYBR Green qPCR Master Mix
with Fluorescein (2X) 31HotStart-IT SYBR Green qPCR Master Mix (2X) 32HotStart-IT SYBR Green qPCR Master Mix with
UDG (2X) 32VeriQuest Probe qPCR Master Mix (2X) 33VeriQuest Probe qPCR Master Mix, No
Reference Dye (2X) 35VeriQuest Fast Probe qPCR Master Mix (2X) 36VeriQuest Fast Probe qPCR Master Mix, No
Reference Dye (2X) 37HotStart-IT Probe qPCR Master Mix (2X) 38HotStart-IT Probe qPCR Master Mix with
UDG (2X) 38
Real-Time Reverse Transcription PCR (qRT-PCR)First-Strand cDNA Synthesis Kit for Real-Time
PCR 39VeriQuest SYBR Green One-Step qRT-PCR
Master Mix (2X) 40VeriQuest SYBR Green One-Step qRT-PCR
Master Mix with Fluorescein (2X) 41VeriQuest Probe One-Step qRT-PCR Master
Mix (2X) 42VeriQuest Probe One-Step qRT-PCR Master
Mix, No Reference Dye (2X) 43HotStart-IT SYBR Green One-Step qRT-PCR
Master Mix Kit 44HotStart-IT Probe One-Step qRT-PCR Master
Mix Kit 44
Reverse Transcription RT-PCR—One-Step Master MixRT-PCR Master Mix (2X) 45FideliTaq RT-PCR Master Mix (2X) 46
Reverse Transcription RT-PCR—KitsFirst-Strand cDNA Synthesis Kit for Real-Time
PCR 47One-Step RT-PCR Kit 47Two-Step RT-PCR Kit 48
PCR PurificationExoSAP-IT® PCR Product Cleanup 49HT ExoSAP-IT High-Throughput PCR Product
Cleanup 50SBE Cleanup Reagent 52PCR Product Pre-Sequencing Kit 52PrepEase® DNA Cleanup Kits 52PrepEase Gel Extraction Kits 52PrepEase PCR Purification 96-Well Plate Kits
(Ultrafiltration) 53
Passive Reference DyesFluorescein Passive Reference Dye 53ROX™ Passive Reference Dye 53
8 888-362-2447 | 216-765-5000 | usb.affymetrix.com
End Point PCR
Standard Taq DNA Polymerase [71160] Taq PCR Master Mix (2X) [71162] RubyTaq DNA Polymerase [71190] RubyTaq PCR Master Mix (2X) [71191]
Long and Accurate PCR FideliTaq DNA Polymerase [71180] FideliTaq PCR Master Mix (2X) [71182] HotStart-IT FideliTaq DNA Polymerase [71155] HotStart-IT FideliTaq PCR Master Mix (2X) [71156]
Hot Start HotStart-IT Taq DNA Polymerase [71195] HotStart-IT Taq PCR Master Mix (2X) [71196] HotStart-IT Binding Protein [71194]
Multiplex Promoter Methylation PCR Kit [MP1100] MagniTaq Multiplex PCR Master Mix (2X) [71199]
PCR Tools | PCR Product Review
Clear Choices for PCR
Real-Time qPCR
Components & Kits First-Strand cDNA Synthesis Kit for Real-Time PCR [75780] VeriQuest Taq DNA Polymerase (2X) [71170]
SYBR VeriQuest SYBR Green qPCR Master Mix (2X) [75600] VeriQuest SYBR Green qPCR Master Mix with Fluorescein
(2X) [75665] VeriQuest Fast SYBR Green qPCR Master Mix (2X) [75690] VeriQuest Fast SYBR Green qPCR Master Mix with
Fluorescein (2X) [75675] HotStart-IT SYBR Green qPCR Master Mix (2X) [75762] HotStart-IT SYBR Green qPCR Master Mix with UDG (2X)
[75760]
Probe VeriQuest Probe qPCR Master Mix (2X) [75650] VeriQuest Probe qPCR Master Mix, No Reference Dye (2X)
[75660] VeriQuest Fast Probe qPCR Master Mix (2X) [75680] VeriQuest Fast Probe qPCR Master Mix, No Reference Dye
(2X) [75685] HotStart-IT Probe qPCR Master Mix (2X) [75766] HotStart-IT Probe qPCR Master Mix with UDG (2X) [75764]
Real-Time Reverse Transcription PCR (qRT-PCR)
One-Step Master Mixes VeriQuest SYBR Green One-Step qRT-PCR Master Mix (2X)
[75705] VeriQuest SYBR Green One-Step qRT-PCR Master Mix with
Fluorescein (2X) [75715] VeriQuest Probe One-Step qRT-PCR Master Mix (2X)
[75700] VeriQuest Probe One-Step qRT-PCR Master Mix, No
Reference Dye (2X) [75710] HotStart-IT SYBR Green One-Step qRT-PCR Master Mix Kit
[75770] HotStart-IT Probe One-Step qRT-PCR Master Mix Kit
[75772]
Kits First-Strand cDNA Synthesis Kit for Real-Time PCR [75780]
Reverse Transcription RT-PCR
One-Step Master Mix RT-PCR Master Mix [2X] [78370] FideliTaq RT-PCR Master Mix [2X] [71185]
Kits One-Step RT-PCR Kit [78350] Two-Step RT-PCR Kit [78355]
PCR Components
Nucleotides 10 mM PCR Nucleotide Mix [77212] 25 mM PCR Nucleotide Mix [77119] 10 mM PCR Nucleotide Mix with dUTP [77330]
Reagents 5M Betaine [77507] Nuclease-Free Water [71786]
9For bulk or alternate pack sizes, email us at [email protected].
PCR Tools | PCR Product Review
PCR Product Selection Guide
End
Point
PCR
Hot St
art P
CR
Long
and
Accur
ate P
CR
Mult
iplex
PCR
Fast
PCR
Trac
king
Dyes P
CR
Mas
ter M
ix Fo
rmat
Stan
d-alo
ne En
zym
e
RT-P
CR
Real-
time P
CR: SY
BR
Real-
time P
CR: Pr
obe
One-S
tep
qRT-
PCR:
SYBR
One-S
tep
qRT-
PCR:
Prob
e
Taq DNA Polymerase [71160] � �Taq PCR Master Mix (2X) [71162] � �RubyTaq™ DNA Polymerase [71190] � � �RubyTaq PCR Master Mix (2X) [71191] � � �FideliTaq™ DNA Polymerase [71180] � � �FideliTaq PCR Master Mix (2X) [71182] � � �VeriQuest® Taq DNA Polymerase [71170] � � �HotStart-IT® Taq DNA Polymerase [71195] � � �HotStart-IT Taq Master Mix (2X) [71196] � � �HotStart-IT FideliTaq DNA Polymerase [71155] � � � �HotStart-IT FideliTaq PCR Master Mix (2X) [71156] � � � �
AMV Reverse Transcriptase [70041] � �M-MLV Reverse Transcriptase [78306] � �Tth DNA Polymerase [70052] � �RT-PCR Master Mix (2X) [78370] � �FideliTaq RT-PCR Master Mix (2X) [71185] � �
First-Strand cDNA Synthesis Kit for Real-Time PCR [75780] � � � �VeriQuest SYBR® Green qPCR Master Mix (2X) [75600] � � �VeriQuest SYBR Green qPCR Master Mix with Fluorescein (2X) [75665] � � �VeriQuest Probe qPCR Master Mix (2X) [75650] � � �VeriQuest Probe qPCR Master Mix, No Reference Dye (2X) [75660] � � �VeriQuest Fast SYBR Green qPCR Master Mix (2X) [75690] � � � �VeriQuest Fast SYBR Green qPCR Master Mix with Fluorescein (2X) [75675] � � � �VeriQuest Fast Probe qPCR Master Mix (2X) [75680] � � � �VeriQuest Fast Probe qPCR Master Mix, No Reference Dye (2X) [75685] � � � �VeriQuest SYBR Green One-Step qRT-PCR Master Mix (2X) [75705] � � �VeriQuest SYBR Green One-Step qRT-PCR Master Mix with Fluorescein (2X) [75715] � � �VeriQuest Probe One-Step qRT-PCR Master Mix (2X) [75700] � � �VeriQuest Probe One-Step qRT-PCR Master Mix, No Reference Dye (2X) [75710] � � �HotStart-IT SYBR Green qPCR Master Mix (2X) [75762] � � �HotStart-IT SYBR Green qPCR Master Mix with UDG (2X) [75760] � � �HotStart-IT Probe qPCR Master Mix (2X) [75766] � � �HotStart-IT Probe qPCR Master Mix with UDG (2X) [75764] � � �
MagniTaq® Multiplex PCR Master Mix [71199] � � � �
Let our simple chart below help you find the optimal reagent for your research needs. Navigate our PCR product portfolio by application to view your options.
10 888-362-2447 | 216-765-5000 | usb.affymetrix.com
PCR Tools | PCR Product Review
Instrument Compatibility with VeriQuest qPCR Master Mixes
Use this chart to determine which of our VeriQuest qPCR master mixes we recommend for your machine. Simply find your thermo cycler across the top row and then locate which of our master mixes are optimized for that instrument.
VeriQuest Products
Enzyme
71170 VeriQuest Taq DNA Polymerase See page 26
SYBR Green Mixes
75600 VeriQuest SYBR Green qPCR Master Mix (2X) See page 27
75665 VeriQuest SYBR Green qPCR Master Mix with Fluorescein (2X) See page 29
Probe Mixes
75650 VeriQuest Probe qPCR Master Mix (2X) See page 33
75660 VeriQuest Probe qPCR Master Mix, No Reference Dye (2X) See page 35
Fast SYBR Green Mixes
75690 VeriQuest Fast SYBR Green qPCR Master Mix (2X) See page 30
75675 VeriQuest Fast SYBR Green qPCR Master Mix with Fluorescein (2X) See page 31
Fast Probe Mixes
75680 VeriQuest Fast Probe qPCR Master Mix (2X) See page 36
75685 VeriQuest Fast Probe qPCR Master Mix, No Reference Dye (2X) See page 37
One-Step qRT-PCR SYBR Green Mixes
75705 VeriQuest SYBR Green One-Step qRT-PCR Master Mix (2X) See page 40
75715 VeriQuest SYBR Green One-Step qRT-PCR Master Mix with Fluorescein (2X) See page 41
One-Step qRT-PCR Probe Mixes
75700 VeriQuest Probe One-Step qRT-PCR Master Mix (2X) See page 42
75710 VeriQuest Probe One-Step qRT-PCR Master Mix, No Reference Dye (2X) See page 43
SYBR Green Mix ABI Pri
sm® 70
00
ABI Pri
sm 77
00
ABI Pri
sm 79
00 HT*
ABI 57
00
ABI 73
00
ABI 75
00*
ABI Ste
pOne
™/St
epOne
Plus™
*
ABI ViiA
™ 7
Bio-Ra
d iCycl
er iQ
® /iQ5
Bio-Ra
d iCycl
er MyiQ
™
Bio-Ra
d CFX
96™
/CFX38
4™
Bio-Ra
d Opti
con2
™
Bio-Ra
d Chro
mo4™
Cephe
id Sm
art Cycl
er®
Corbett
Rotor
-Gen
e™
Eppe
ndorf
Mast
ercycl
er® ep
realp
lex
Fluidi
gm Bi
oMark
™
Illum
ina Ec
o™*/H
elixis
Pixo
™
Qiagen
Rotor
-Gen
e™ Q
*
Roch
e Ligh
tCycler
® 1.0,
1.5, 2
.0*
Roch
e Ligh
tCycler
480/1
536*
Strata
gene
Mx3
000P
™*
Strata
gene
Mx3
005P
™*
Strata
gene
Mx4
000™
TaKaR
a TP-8
00™
VeriQuest® SYBR® Green qPCR Master Mix (2X) [75600]
VeriQuest SYBR Green qPCR Master Mix with Fluorescein (2X) [75665]
Probe Mix
VeriQuest Probe qPCR Master Mix (2X) [75650]
VeriQuest Probe qPCR Master Mix, No Reference Dye (2X) [75660]
Fast SYBR Green Mix
VeriQuest Fast SYBR Green qPCR Master Mix (2X) [75690]
VeriQuest Fast SYBR Green qPCR Master Mix with Fluorescein (2X) [75675]
Fast Probe Mix
VeriQuest Fast Probe qPCR Master Mix (2X) [75680]
VeriQuest Fast Probe qPCR Master Mix, No Reference Dye (2X) [75685]
One-Step qRT-PCR SYBR Green Mix
VeriQuest SYBR Green One-Step qRT-PCR Master Mix (2X) [75705]
VeriQuest SYBR Green One-Step qRT-PCR Master Mix with Fluorescein (2X) [75715]
One-Step qRT-PCR Probe Mix
VeriQuest Probe One-Step qRT-PCR Master Mix (2X) [75700]
VeriQuest Probe One-Step qRT-PCR Master Mix, No Reference Dye (2X) [75710]
* Instruments can be set to fast mode cycling when using fast mode master mixes. Indicates preferred kit for this cycler
Indicates maybe also used for this cycler
11For bulk or alternate pack sizes, email us at [email protected].
PCR Tools | End-Point PCR
71160 50 units 250 units 1,000 units 5,000 unitsCustoms by request
Source:E. coli strain expressing a full length, unmodified clone of Taq DNA Polymerase from Thermus aquaticus(1-3).
Description:Taq DNA Polymerase is a thermostable enzyme which has a highly processive 5'→3' polymerase activity and a 5'→3' exonuclease activity (1). Taq enzyme is functionally tested and has no detectable contaminating endonucleases. Taq DNA Polymerase withstands repeated incubations at 95°C without a significant decrease in enzyme activity, and is suitable for routine PCR. Taq DNA Polymerase is sup-plied with 10X PCR Reaction Buffer plus a separate tube of 25 mM MgCl2 for optimizing PCR conditions. Each lot is tested for product yield and length in PCR amplification.
Applications:�� Routine PCR amplification�� Suitable for real-time qPCR
Properties:Activator: Mg2+
Purity:Free from detectable non-specific nucleases.
Storage buffer:20 mM Tris-HCl, pH 8.5, 1 mM DTT, 0.1 mM EDTA, 100 mM KCI, 50% glycerol, stabilizers.
Unit definition:One unit incorporates 10 nmol of total nucleotides into acid-insoluble material in 30 minutes at 74°C in a total volume of 50 µl.
Concentration:5 units/µl
Assay conditions:The reaction mixture (50 µl) contains 25 mM TAPS, pH 9.3 (at 25°C), 50 mM KCl, 2 mM MgCl2, 1 mM 2-mercaptoethanol, 200 µM dNTPs, 250 µg/ml activated salmon sperm DNA, and Taq DNA Polym-erase. After incubation at 74°C for 10 minutes, acid insoluble material is determined.
Functional test:Tested in a standard PCR.
Functionally tested 10X PCR Reaction Buffer (included):100 mM Tris-HCl (pH 8.6), 500 mM KCl, 15 mM MgCl2Functionally tested MgCl2 (included):25 mM solution
Shipping and storage:Shipped on dry ice. Store at -20°C.
References:1. Innis, M. A., Myambo, K. B., Gelfand, D. H. and
Brow, M. A. (1988) Proc. Natl. Acad. Sci. 85, 9436-9440.
2. Lawyer, F. C., Stoffel, S., Saiki, R. K., Myambo, K., Drummond, R. and Gelfand, D. H. (1989) J. Biol. Chem. 264, 6427-6437.
3. Innis, M. A. and Gefland, D. H. (1990) PCR Protocols: A Guide to Methods and Applications, Academic Press.
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Agarose - LE32802 25 gm 100 gm 250 gm 500 gm
ExoSAP-IT PCR Product Cleanup78250 20 reactions78200 100 reactions78201 500 reactions78202 2,000 reactions78205 5,000 reactions
PCR Markers, 50 - 2000 bp76710 250 µl
PCR Nucleotide Mix, 10 mM Solution(10 mM each dATP, dCTP, dGTP, dTTP)77212 500 µl
PCR Nucleotide Mix, 25 mM Solution77119 500 µl
PCR Nucleotide Mix with dUTP, 10 mM Solution77330 500 µl
Water, Nuclease-Free71786 10 x 1 ml 100 ml 500 ml 1 L 5 L
Taq PCR Kit71161 100 reactions
Taq DNA Polymerase
Fig. 1. DNA fragments ranging from 1 kb to 6 kb amplified from λ DNA.
0.5 kb –
1 kb –
2 kb –
5 kb –
12 kb –
M 1 2 3 4 5 6 kbp
12 888-362-2447 | 216-765-5000 | usb.affymetrix.com
71162 100 reactionsCustoms by request
Taq PCR Master Mix is supplied as a convenient 2X pre-mixed formulation containing Taq DNA Polymerase, Ultrapure nucleotides, and reaction buffer optimized for a wide variety of PCR applica-tions. Taq PCR Master Mix provides robust and reliable performance in PCR amplification. Since the mix is prepared from high quality reagents and is thoroughly QC tested, experimental variability is significantly reduced.
Applications:�� Routine PCR amplification�� Suitable for real-time qPCR�� High-throughput PCR
ConvenientThe mix saves time and reduces potential contami-nation errors by eliminating several pipetting steps. For a 50 µl reaction, simply add 25 µl of Taq PCR Master Mix to primers, DNA template, and PCR-Qualified H2O.
StableThe mix withstands repeated freeze-thaw cycles with no observed decrease in performance (Fig. 1). It is also stable at 4°C for extended periods of time. In fact, rigorous stability studies have been done at Affymetrix showing the mix to be stable at room temperature (Fig. 2).
ScaleableThe end-user may set-up 10 µl, 25 µl, 50 µl, or 100 µl reactions.
Ideal for high-throughputTaq PCR Master Mix is ideal for high-throughput ap-plications such as screening transformants by colony PCR (Fig. 3) or transgenic studies.
Sensitive and robustThe mix may be used to amplify PCR products from low-copy genomic DNA targets as well as low to moderately abundant cDNAs (Figs. 1-3). Excellent results are also obtained with long PCR products (Fig. 4).
Functional test:Functionally tested in PCR according to the standard Affymetrix PCR protocol. This protocol is based upon the amplification of a 500 bp DNA sequence.
Taq PCR Master Mix Formulation (2X):20 mM Tris-HCl (pH 8.6), 100 mM KCl, 3 mM MgCl2, 0.4 mM dNTPs (dATP, dCTP, dGTP, dTTP), 50 units/ml Taq DNA Polymerase, stabilizers.
Components:4 x 625 µl (100 x 50 µl reactions or 200 x 25 µl reactions)
Shipping and storage:Shipped on dry ice. Store at -20°C. Mix well prior to use.
PCR Tools | End-Point PCR
Taq PCR Master Mix (2X)
Fig. 1. Freeze-thaw stability of Taq PCR Master Mix. Amplification of a segment of the single-copy numb gene from 50 pg human genomic DNA. This represents about 30 total target copies. Following 25 freeze-thaw cycles, no significant effect on performance was observed.
Fig. 2. Elevated temperature stability. Taq PCR Master Mix was stored at 4°C and room temper-ature for two months and its performance was compared against a freshly prepared mix. Segments of the single-copy p53 gene (left) and numb gene (right) were amplified from 50 pg human genomic DNA. (This represents about 30 total target copies.) Under these storage conditions, PCR amplifica-tion was not significantly affected.
Fig. 3. Colony PCR. Amplification of a segment of the T4 DNA Polymerase gene overexpressed in several different E. coli M5248 colonies.
Fig. 4. Amplification of targets from genomic DNA. Segments of the single-copy NRAGE gene (a) and numb gene (b) were amplified from 50 pg and 50 ng of human genomic DNA respectively.
M 0 15 25Freeze thaws
0.4 kb –
1.0 kb –
← numb (455 bp)
M Fresh 4°C RT M Fresh 4°C RT
p53 (247 bp) →
0.5 kb –
0.1 kb –
← numb (455 bp)
M
← 574 bp1.0 kb –
0.5 kb –
M
NRAGE (1369 bp) →
a
2.0 kb –
1.0 kb –
← numb (4.6 kb)
M
b
5.0 kb –
3.0 kb –
71161 100 reactions (100 µl/rctn)
The Taq PCR Kit includes the necessary reagents to amplify DNA from a wide variety of templates. All components, including Ultrapure PCR Qualified Water and Ultrapure PCR Nucleotide Mix, are
functionally tested in standard PCR to ensure reliable results. The kit also includes two PCR buffers for standard reactions, one with and one without MgCl2, and a separate tube of 25 mM magnesium chloride for optimizing PCR conditions.
Kit components:Taq DNA Polymerase, 250 units10 mM PCR Nucleotide Mix, Ultrapure10X PCR Buffer with MgCl210X PCR Buffer without MgCl225 mM MgCl2PCR-Qualified Water, Ultrapure
Taq PCR Kit
13For bulk or alternate pack sizes, email us at [email protected].
PCR Tools | End-Point PCR
71190 50 units 250 units 1,000 units 5,000 unitsCustoms by request
Source:E. coli strain expressing a clone of Taq DNA Polym-erase from Thermus aquaticus(1-3).
Description:RubyTaq DNA Polymerase contains high-quality Taq DNA Polymerase with two inert dyes that serve as tracking dyes in gel electrophoresis. This greatly simplifies analysis of PCR reactions because no load-ing dyes or compounds that increase sample density need to be added prior to gel electrophoresis. During gel electrophoresis, RubyTaq Polymerase separates into 2 colors, magenta and yellow, for easy visualiza-tion (Fig. 1). The magenta dye runs in a range be-tween 500 bp (2% gels) and 1,500 bp (0.8% gels), and the yellow dye runs at less than 10 bp.
Applications:�� Routine PCR amplification�� TA-vector cloning�� Amplification prior to in vitro transcription
Easy-to-see, easy-to-load�� Load directly to gel from thermocycler�� No additional loading buffer needed�� RubyTaq separates into 2 colors – magenta and yellow – during gel electrophoresis.
Because the dyes are inert during PCR, RubyTaq DNA Polymerase provides the same outstanding performance as standard Taq DNA Polymerase (Fig. 2). In addition, the dyes do not affect the migration rates of DNA and have no effect on the following procedures:�� PCR with co-solvents (e.g., betaine and DMSO)�� Restriction digestion�� Automated or manual DNA sequencing�� Ligation-based or topoisomerase-based (TOPO®) TA cloning�� ExoSAP-IT PCR Cleanup
If the dyes must be removed prior to another ap-plication (e.g., in vitro transcription), use phenol/chloroform extraction and ethanol precipitation, or the PrepEase Sequencing Dye Cleanup Kit (PN 78776/77).
Properties:Activator: Mg2+
Purity:Free from detectable non-specific nucleases.
Storage buffer:20 mM Tris-HCl, pH 8.5, 1 mM DTT, 0.1 mM EDTA, 50 mM KCI, 50% glycerol, inert dyes, and stabilizers.
Unit definition:One unit incorporates 10 nmol of total nucleotides into acid-insoluble material in 30 minutes at 74°C in a total volume of 50 µl.
Concentration:1 unit/µl
Assay conditions:The reaction mixture (50 µl) contains 25 mM TAPS, pH 9.3 (at 25°C), 50 mM KCl, 2 mM MgCl2, 1 mM 2-mercaptoethanol, 200 µM dNTPs, 250 µg/ml acti-vated salmon sperm DNA, and cloned RubyTaq DNA Polymerase. After incubation at 74°C for 10 minutes, acid-insoluble material is determined.
Functional test:Functionally tested for PCR.
Functionally tested 10X PCR Reaction Buffer without MgCl2 (included):100 mM Tris-HCl (pH 8.6) and 500 mM KCl
Functionally tested MgCl2 (included):25 mM solution
Shipping and storage:Shipped on dry ice. Store at -20°C.
References:1. Innis, M. A., Myambo, K. B., Gelfand, D. H. and
Brow, M. A. (1988) Proc. Natl. Acad. Sci. 85, 9436-9440.
2. Lawyer, F. C., Stoffel, S., Saiki, R. K., Myambo, K., Drummond, R. and Gelfand, D. H. (1989) J. Biol. Chem. 264, 6427-6437.
3. Innis, M. A. and Gefland, D. H. (1990) PCR Protocols: A Guide to Methods and Applications, Academic Press.
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Agarose - Separation ≥ 500 bpGenetic Performance Certified™75817 25 gm 100 gm 250 gm 500 gm
ExoSAP-IT PCR Product Cleanup78250 20 reactions78200 100 reactions78201 500 reactions78202 2,000 reactions78205 5,000 reactions
PCR Nucleotide Mix, 10 mM Solution(10 mM each dATP, dCTP, dGTP, dTTP)77212 500 µl
TAE Buffer, 10X Solution75904 1 L 5 L
TBE Buffer, 5X Solution75891 1 L 5 L
Water, Nuclease-Free71786 10 x 1 ml 100 ml 500 ml 1 L 5 L
RubyTaq DNA Polymerase
Fig. 2. Performance of RubyTaq DNA Polymerase versus Taq DNA Polymerase. Single-copy targets were amplified from the indi-cated amounts of human genomic DNA. For the 455 bp numb ampli-con, 50 pg of template represents about 30 total target copies in the reaction. T is Taq DNA Polymerase, R is RubyTaq DNA Polymerase, and M is the DNA marker.
M T R T R T R
5.0 kb – ← 4.6 kb numb (50 ng)← 1.55 kb NRAGE (10 ng)
← 455 bp numb (50 pg)
1.0 kb –
100 bp –
Fig. 1. RubyTaq DNA Polymerase visualization during gel electrophoresis. RubyTaq Polymerase in PCR does not require the use of additional loading buffer. Simply load onto an agarose gel directly after cycling. During electrophoresis, RubyTaq separates into 2 colors, magenta (runs between 500 bp [2% gels] and 1,500 bp [0.8% gels]) and yellow (runs less than 10 bp). The indicated volumes of RubyTaq DNA Polymerase were run on a 1% TAE agarose gel.
14 888-362-2447 | 216-765-5000 | usb.affymetrix.com
71191 100 reactions 500 reactionsCustoms by request
RubyTaq PCR Master Mix is supplied as a convenient 2X pre-mixed formulation containing Taq DNA Polymerase, Ultrapure nucleotides, reaction buffer, and two inert dyes optimized for a wide variety of PCR applications. RubyTaq enzyme greatly simplifies analysis of PCR reactions because no loading dyes or compounds that increase sample density need to be added prior to gel electrophoresis. During gel elec-trophoresis, RubyTaq PCR Master Mix separates into 2 colors, magenta and yellow, for easy visualization. The magenta dye runs in a range between 500 bp (2% gels) and 1,500 bp (0.8% gels) and the yellow dye runs at less than 10 bp.
RubyTaq PCR Master Mix has been designed to streamline high-throughput PCR applications and provides for robust and reliable performance. Since the mix is prepared from high quality reagents and is thoroughly QC tested, experimental variability is significantly reduced.
Applications:�� Routine PCR amplification�� TA vector cloning�� Amplification prior to in vitro transcription�� High-throughput PCR
Easy-to-see, easy-to-load�� Load directly to gel from thermocycler�� No additional loading buffer needed�� RubyTaq separates into 2 colors – magenta and yellow – during gel electrophoresis.
Ultimate convenienceThe mix saves time and reduces potential contami-nation errors by eliminating several pipetting steps. For a 50 µl reaction, simply add 25 µl of RubyTaq PCR Master Mix to primers, DNA template, and PCR-Qualified H2O. Reactions can be easily performed in 10 µl, 25 µl, 50 µl, or 100 µl volumes. In addition, PCR reactions are loaded directly onto agarose gels without adding any loading dyes or buffers.
StableThe mix withstands repeated freeze-thaw cycles and extended periods at room temperature with no observed decrease in performance (Fig. 1).
Ideal for high-throughputRubyTaq PCR Master Mix is ideal for high-throughput applications such as:�� Screening transformants by colony PCR�� Transgenic genotyping�� Ligation-based or topoisomerase-based (TOPO) TA cloning
The mix may be used to amplify low-copy targets from complex templates and can also amplify a variety of product lengths.
Functional test:Functionally tested for PCR according to the stan-dard Affymetrix PCR protocol. This protocol is based upon the amplification of a 500 bp DNA sequence.
RubyTaq PCR Master Mix Formulation (2X):20 mM Tris-HCl (pH 8.6), 100 mM KCl, 4 mM MgCl2, 0.4 mM dNTPs (dATP, dCTP, dGTP, dTTP), 80 units/ml Taq DNA Polymerase, inert dyes, stabilizers.
Components:4 x 625 µl RubyTaq PCR Master Mix, 2X
(100 x 50 µl reactions or 200 x 25 µl reactions)
Shipping and storage:Shipped on dry ice. Store at -20°C. Mix well prior to use.
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ExoSAP-IT PCR Product Cleanup78250 20 reactions78200 100 reactions78201 500 reactions78202 2,000 reactions78205 5,000 reactions
PCR Markers, 50 - 2000 bp76710 250 µl
PCR Nucleotide Mix, 10 mM Solution(10 mM each dATP, dCTP, dGTP, dTTP)77212 500 µl
PrepEase Gel Extraction Kit78756 50 preps78757 250 preps
TAE Buffer, 10X Solution75904 1 L 5 L
TBE Buffer, 5X Solution75891 1 L 5 L
Water, Nuclease-Free71786 10 x 1 ml 100 ml 500 ml 1 L 5 L
RubyTaq PCR Master Mix (2X)
PCR Tools | End-Point PCR
M
← 455 bp numb (50 pg)1.0 kb –
100 bp –
-20°CF/T RT
Fig. 1. Stability of RubyTaq PCR Master Mix. RubyTaq PCR Master Mix stability was assessed by comparing a reference mix stored at -20°C versus a sample subjected to 15 freeze-thaw cycles (F/T) and a sample stored at room temperature for one month (RT). The single-copy numb gene was amplified from 50 pg of human genomic DNA which represents about 30 total target copies in the reaction. Under these test conditions, performance of the master mix is not significantly affected.
15For bulk or alternate pack sizes, email us at [email protected].
71180 50 units 250 units 1,000 units 5 x 250 units 5,000 unitsCustoms by request
Source:Recombinant proteins expressed in E. coli.
Description:FideliTaq DNA Polymerase combines high quality recombinant Taq DNA Polymerase with a high-fidelity, proofreading polymerase. This enzyme blend has the 5'→3' exonuclease activity of Taq DNA Polymerase as well as the 3'→5' exonuclease activity of the proofreading polymerase. FideliTaq DNA Polymerase increases amplification fidelity up to 6 times over Taq DNA Polymerase alone and allows for amplification of longer product sizes(1-4) (Fig. 1). The enzyme blend generates PCR products whose ends are compatible with either blunt-end or TA cloning procedures(5) with A-tailed ends favored over blunt ends in an approximately 3 to 1 ratio.
Applications:�� PCR-mediated cloning�� Long and accurate PCR amplification
Properties:Activator: Mg2+
Purity:Free of detectable non-specific nucleases.
Storage buffer:20 mM Tris-HCl, pH 8.5, 1 mM DTT, 0.1 mM EDTA, 100 mM KCI, 50% glycerol, stabilizers.
Unit definition:One unit incorporates 10 nmol of total nucleotides into acid-insoluble material in 30 minutes at 74°C in a total volume of 50 µl(6-7).
Concentration:5 units/µl
Assay conditions:The reaction mixture contains 25 mM TAPS, pH 9.3 (at 25°C), 50 mM KCl, 2 mM MgCl2, 1 mM β-ME, 200 µM dNTPs, 250 µg/ml activated salmon sperm DNA, and FideliTaq DNA Polymerase. Following incubation at 74°C for 10 minutes, acid-insoluble material is determined.
Functional test:Functionally tested in PCR to amplify a 20.7 kb product.
Functionally tested 10X PCR Reaction Buffer (included):100 mM Tris-HCl (pH 8.6), 500 mM KCl, 15 mM MgCl2Functionally tested MgCl2 (included):25 mM solution
Shipping and storage:Shipped on dry ice. Store at -20°C.
References:1. Barnes, W. M. (1994) Proc. Natl. Acad. Sci. USA
91, 2216-2220.2. Cheng, S., Fockler, C., Barnes, W. M., and
Higuchi, R. (1994) Proc. Natl. Acad. Sci. USA 91, 5695-5699.
3. Barnes, W. M. (1992) Gene 112, 29-35.4. Cline, J., Braman, J. C., and Hogrefe, H. H. (1996)
Nucl. Acids Res. 24, 3546-3551.5. Magnuson, V. L., Ally, D. S., Nylund, S. J.,
Karanjawala, Z. E., Rayman, J. B., Knapp, J. I., Lowe, A. L., Ghosh, S., and Collins, F. S. (1996) BioTechniques 4, 700-709.
6. Innis, M. A., Myambo, K. B., Gelfand, D. H. and Brow, M. A. (1988) Proc. Natl. Acad. Sci. 85, 9436-9440.
7. Lawyer, F. C., Stoffel, S., Saiki, R. K., Myambo, K., Drummond, R. and Gelfand, D. H. (1989) J. Biol. Chem. 264, 6427-6437.
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HT ExoSAP-IT High-Throughput PCR Product Cleanup78395 480 rctns x 8-tube strip 5,760 rctns x 1 plate (12 x 8-tube strips) 23,040 rctns - 4 plates 1,000 rctns (2 ml) 5,000 rctns (10 ml)
PCR Markers, 50 - 2000 bp76710 250 µl
PCR Nucleotide Mix, 10 mM Solution(10 mM each dATP, dCTP, dGTP, dTTP)77212 500 µl
Water, Nuclease-Free71786 10 x 1 ml 100 ml 500 ml 1 L 5 L
FideliTaq DNA Polymerase
PCR Tools | Long and Accurate PCR
Fig. 1. Amplification of diverse targets by FideliTaq DNA Polymerase. Target (source): Lane 1: Numb, 455 bp (10 ng human genomic DNA); Lane 2: NRAGE, 1.55 kb (10 ng human genomic DNA); Lane 3: Numb, 4.6 kb (10 ng human genomic DNA); Lane 4: Lambda target, 20.7 kb (10 ng lambda DNA). A total of 25 cycles was used for lambda target and 35 cycles for human genomic DNA targets. A final concentration of 1.5 mM MgCl2 was used in all reactions.
M 1 2 3 4
12.0 kb –
3.0 kb –
1.65 kb –
500 bp –
16 888-362-2447 | 216-765-5000 | usb.affymetrix.com
PCR Tools | Long and Accurate PCR
71182 100 reactionsCustoms by request
FideliTaq PCR Master Mix (2X) combines Taq DNA Polymerase, a high-fidelity, proofreading polym-erase and Ultrapure nucleotides in a proprietary buffer formulation. This ready-to-use mix increases amplification fidelity up to 6 times over Taq DNA Polymerase alone and allows for amplification of longer product sizes(1-4). The enzyme blend generates PCR products whose ends are compatible with either blunt-end or TA cloning procedures(5) with A-tailed ends favored over blunt ends in an approximately 3 to 1 ratio. FideliTaq PCR Master Mix provides robust and reliable performance for PCR applications in which high-fidelity or longer product sizes are re-quired. Since the mix is pre-formulated, experimental variability is significantly reduced.
Applications:�� PCR-mediated cloning�� Long and accurate PCR amplification�� High-throughput PCR
ConvenientThe mix saves time and reduces potential contami-nation errors by eliminating several pipetting steps. For a 50 µl reaction, simply add 25 µl of FideliTaq PCR Master Mix to primers, DNA template, and PCR-Qualified H2O. Reactions can be easily performed in 10 µl, 25 µl, 50 µl or 100 µl volumes.
High fidelityFideliTaq DNA Polymerase gives up to 6-fold higher fidelity than Taq DNA Polymerase, ideal for cloning and microarray applications.
Increase product size and yieldAmplify short and long PCR products from complex DNA templates with little or no optimization (Fig. 1). For PCR products greater than 2 kb, yields are greatly increased with FideliTaq DNA Polymerase and an enhanced buffer system.
Improve specificity and sensitivityAmplify PCR products with low background and from low-copy targets, essential for demanding genomic and cDNA applications with limited sample material (Fig. 2).
StableThe mix withstands repeated freeze-thaw cycles with no observed decrease in performance (Fig. 3).
Functional test:Functionally tested in PCR to amplify a 20.7 kb product.
FideliTaq PCR Master Mix Formulation (2X):50 mM Tris, pH 8.8, 100 mM KCl, 3 mM MgCl2, 0.4 mM dNTPs (dATP, dCTP, dGTP, dTTP), 50 units/ml FideliTaq DNA Polymerase, stabilizers.
Components:4 x 625 µl FideliTaq PCR Master Mix, 2X
(100 x 50 µl reactions or 200 x 25 µl reactions)
Shipping and storage:Shipped on dry ice. Store at -20°C. Mix well prior to use.
References:1. Barnes, W. M. (1994) Proc. Natl. Acad. Sci. USA
91, 2216-2220.2. Cheng, S., Fockler, C., Barnes, W. M., and
Higuchi, R. (1994) Proc. Natl. Acad. Sci. USA 91, 5695-5699.
3. Barnes, W. M. (1992) Gene 112, 29-35.4. Cline, J., Braman, J. C., and Hogrefe, H. H. (1996)
Nucl. Acids Res. 24, 3546-3551.5. Magnuson, V. L., Ally, D. S., Nylund, S. J.,
Karanjawala, Z. E., Rayman, J. B., Knapp, J. I., Lowe, A. L., Ghosh, S., and Collins, F. S. (1996) BioTechniques 4, 700-709.
FideliTaq PCR Master Mix (2X)
Fig. 1. Range of targets amplified with FideliTaq PCR Master Mix. Single-copy NRAGE, 1.55 kb (Lane 1) and Numb, 7.7 kb (Lane 2) were amplified from 1 ng human genomic DNA. Single-copy β-globin, 23.0 kb (Lane 3) was amplified from 100 ng human genomic DNA. Both the 20.7 kb (Lane 4) and 35.0 kb (Lane 5) lambda targets were amplified from 1 ng lambda DNA. No magnesium optimization was required, as 1.5 mM final magnesium concentration was used in all reactions.
M 1 2 3 4 5
12.0 kb –
7.0 kb –
1.65 kb –
500 bp –
Fig. 2. Sensitivity of FideliTaq PCR Master Mix. The single-copy Numb gene was amplified from the indicated amounts of human genomic DNA. Approximately one mammalian cell is represented by 3 pg of genomic DNA.
M 300 30 3 pg
650 bp –500 bp –400 bp –
Fig. 3. Freeze-thaw stability of FideliTaq PCR Master Mix. The master mix was subjected to 15 freeze-thaw cycles alternating between dry ice and room temperature. Following freeze-thaw cycles (F/T), the mix was compared to control mix (C) before treatment. Both short and long targets are shown to demonstrate the robust nature of the mix. Single-copy Numb, 455 bp (Lane 1) was amplified from 30 pg human genomic DNA. β-globin 23 kb target (Lane 2) was amplified from 100 ng human genomic DNA. Lambda 35 kb target (Lane 3) was amplified from 1 ng lambda DNA.
1 1 2 2 3 3M C F/T C F/T C F/T
12.0 kb –
500 bp –
17For bulk or alternate pack sizes, email us at [email protected].
PCR Tools | Long and Accurate PCR
71155 50 units 250 units 1,000 units 5,000 unitsCustoms by request
Source:All proteins are recombinant versions expressed in E. coli.
Description:HotStart-IT FideliTaq DNA Polymerase combines a novel hot start method developed at Affymetrix with the long and accurate amplification properties of FideliTaq DNA Polymerase. The hot start method, called primer sequestration, uses a binding protein to reduce or eliminate non-specific primer-extension products which may be generated at lower tempera-tures during assembly of PCR reactions. Following the initial denaturation step during PCR, the binding protein is inactivated and the primers are free to participate in the amplification reaction. This novel method enhances many complex PCR reactions by increasing specificity and yield (Fig. 1) as well as sensitivity (Fig. 2).
FideliTaq DNA Polymerase combines Taq DNA Polym-erase with a high-fidelity, proofreading polymerase. This enzyme blend has the 5'→3' exonuclease activity of Taq DNA Polymerase as well as the 3'→5' exonuclease activity of the proofreading enzyme. FideliTaq DNA Polymerase increases fidelity up to 6-fold over Taq DNA Polymerase alone and allows for amplification of longer products(1-4) (Fig. 3). The enzyme blend generates PCR products whose ends are compatible with either blunt-end or TA cloning procedures(5), with A-tailed ends favored over blunt ends in an approximately 3 to 1 ratio.
Applications:�� High-specificity and high-sensitivity PCR amplification�� Extremely long PCR amplification�� PCR-mediated cloning
Advantages:�� Room temperature reaction set-up.�� Minimizes amplification of non-specific products and primer-dimers (Fig. 1).�� High specificity and sensitivity (Fig. 2)�� Generates long PCR amplicons (Fig. 3)�� Up to 6-fold higher fidelity than Taq polymerase
Properties:Activator: Mg2+
Purity:Free from detectable non-specific nucleases.
Storage buffer:20 mM Tris-HCl (pH 8.5), 1 mM DTT, 0.1 mM EDTA, 200 mM KCI, 50% glycerol, and stabilizers.
Unit definition:One unit incorporates 10 nmol of total nucleotides into acid-insoluble material in 30 minutes at 74°C in a total volume of 50 µl(6-7).
Concentration:2.5 units/µl
Assay conditions:The reaction mixture contains 25 mM TAPS, pH 9.3 (at 25°C), 50 mM KCl, 2 mM MgCl2, 1 mM 2-mercaptoethanol, 200 µM dNTPs, 250 µg/ml activated salmon sperm DNA, and Taq DNA Polymerase. Following incubation at 74°C for 10 minutes, acid-insoluble material is determined.
Functional tests:PCR with HotStart-IT FideliTaq DNA Polymerase shifts production of primer-dimers to a specific target of 306 bp from 1 ng of human genomic DNA rela-tive to FideliTaq DNA Polymerase.
PCR with HotStart-IT FideliTaq DNA Polymerase generates a 20.7 kb product.
Functionally tested 10X PCR Reaction Buffer (included): 100 mM Tris-HCl (pH 8.6), 500 mM KCl, 15 mM MgCl2Functionally tested MgCl2 (included):25 mM solution
Shipping and storage:Shipped on dry ice. Store at -20°C.
References:1. Barnes, W. M. (1994) Proc. Natl. Acad. Sci. USA
91, 2216-2220. 2. Cheng, S., Fockler, C., Barnes, W. M., and
Higuchi, R. (1994) Proc. Natl. Acad. Sci. USA 91, 5695-5699.
3. Barnes, W. M. (1992) Gene 112, 29-35. 4. Cline, J., Braman, J. C., and Hogrefe, H. H. (1996)
Nucl. Acids Res. 24, 3546-3551. 5. Magnuson, V. L., Ally, D. S., Nylund, S. J.,
Karanjawala, Z. E., Rayman, J. B., Knapp, J. I., Lowe, A. L., Ghosh, S., and Collins, F. S. (1996) BioTechniques 4, 700-709.
6. Innis, M. A., Myambo, K. B., Gelfand, D. H. and Brow, M. A. (1988) Proc. Natl. Acad. Sci. 85, 9436-9440.
7. Lawyer, F. C., Stoffel, S., Saiki, R. K., Myambo, K., Drummond, R. and Gelfand, D. H. (1989) J. Biol. Chem. 264, 6427-6437.
HotStart-IT FideliTaq DNA Polymerase
Fig. 3. Extreme amplicon length using HotStart-IT FideliTaq DNA Polymerase. Two long PCR products of 20.7 kb and 35 kb were amplified from lambda DNA using 1 ng and 10 ng of DNA respectively. Cycling conditions consisted of a short initial denatur-ation phase at 95°C for 20 seconds and 25 cycles of 95°C for 5 sec-onds and 68°C for 20 minutes.
Fig. 2. Sensitivity of HotStart-IT FideliTaq DNA Polymerase. A 455 bp fragment of the single-copy numb gene was amplified from the indicated amounts of human genomic DNA. The polymerase is extremely sensitive as amplification can be achieved from approxi-mately one human cell.
500 bp –
100 bp –
3 pg (1 cell)
30 pg
– numb, 455 bp
300 pg
500 bp –
100 bp –
no HotStart
HotStart
– numb, 306 bp
– primer-dimers
12 kb –
5 kb –
20.7 kb35 kb
Fig. 1. Increased specificity of HotStart-IT FideliTaq DNA Polymerase. A 306 bp fragment of the single-copy numb gene was amplified from 1 ng of human genomic DNA with and without HotStart-IT technology. The primers in this assay were designed with 3 bases of overlap at their 3'-ends to favor primer-dimer formation during reaction set-up at room temperature. Results demonstrate a shift from mainly primer-dimers to the desired product when HotStart-IT is used.
18 888-362-2447 | 216-765-5000 | usb.affymetrix.com
71156 25 reactions 100 reactions 500 reactionsCustoms by request
HotStart-IT FideliTaq Master Mix combines a novel hot start method developed at Affymetrix with the long and accurate amplification properties of FideliTaq DNA Polymerase. The hot start method, called primer sequestration, uses a binding protein to reduce or eliminate non-specific primer-extension products which may be generated at lower temperatures during assembly of PCR reactions. Following the initial denaturation step during PCR, the binding protein is inactivated and the primers are free to participate in the amplification reaction. This novel method enhances many complex PCR reactions by increasing specificity and yield (Fig. 1) as well as sensitivity (Fig. 2).
HotStart-IT FideliTaq Master Mix combines HotStart-IT FideliTaq DNA Polymerase and Ultrapure nucleotides in a proprietary reaction buffer. Simply add template, primers, and water, and the reactions are ready for cycling. The mix increases amplification fidelity up to 6-fold over Taq DNA Polymerase alone and generates much longer PCR products(1-4) (Fig. 3). PCR products have ends which are compatible with either blunt-end or TA cloning procedures(5) with A-tailed ends favored over blunt ends in an approximately 3 to 1 ratio.
Applications:�� High-specificity and high-sensitivity PCR amplification�� Extremely long PCR amplification�� PCR-mediated cloning�� High-throughput PCR
ConvenientSave time and reduce potential contamination errors by eliminating several pipetting steps. For a 50 µl reaction, simply add 25 µl of HotStart-IT FideliTaq Master Mix to primers, DNA template, and PCR-Qualified H2O. Reactions can be tailored from 10 µl to 100 µl volumes.
Room temperature reaction assembly is possible because of the hot start feature.
Novel hot start technologySince the polymerase is not chemically-inactivated, no extensive heat-activation step is necessary which reduces damage to precious DNA samples.
Higher specificity and sensitivityMinimize amplification of non-specific products and primer-dimers (Fig. 1). Amplify PCR products with low background and from low-copy targets, essential for demanding genomic and cDNA applications with limited sample material (Fig. 2).
High fidelityObtain up to 6-fold higher fidelity than Taq DNA Polymerase, ideal for cloning and microarray applications.
Increase product size and yieldAmplify very long PCR products from complex DNA templates with little or no optimization (Fig. 3). For PCR products greater than 2 kb, yields are greatly increased.
StableRepeated freeze-thaw cycles have no observed effect on performance (Figs. 1 and 3).
Functional tests:PCR with HotStart-IT FideliTaq Master Mix shifts production of primer-dimers to a specific target of 306 bp from 1 ng of human genomic DNA relative to FideliTaq Master Mix.
PCR with HotStart-IT FideliTaq Master Mix generates a 20.7 kb product.
HotStart-IT FideliTaq Master Mix Formulation (2X):HotStart-IT FideliTaq Master Mix combines FideliTaq DNA Polymerase with a recombinant hot start protein in a unique buffer formulation. Magnesium and nucleotide concentrations are 3 mM and 0.4 mM each, respectively.
Components:25 reactions 625 µl100 reactions 4 x 625 µl500 reactions 12.5 ml
Shipping and storage:Shipped on dry ice. Store at -20°C. Mix well prior to use.
References:1. Barnes, W. M. (1994) Proc. Natl. Acad. Sci. USA
91, 2216-2220.2. Cheng, S., Focklet, C., Barnes, W. M., and
Higuchi, R. (1994) Proc. Natl. Acad. Sci. USA 91, 5695-5699.
3. Barnes, W. M. (1992) Gene 112, 29-35.4. Cline, J., Braman, J. C., and Hogrefe, H. H. (1996)
Nucl. Acids Res. 24, 3546-3551.5. Magnuson, V. L., Ally, D. S., Nylund, S. J.,
Karanjawala, Z. E., Rayman, J. B., Knapp, J. I., Lowe, A. L., Ghosh, S., and Collins, F. S. (1996) BioTechniques 4, 700-709.
PCR Tools | Long and Accurate PCR
HotStart-IT FideliTaq PCR Master Mix (2X)
Fig. 1. Increased specificity of the HotStart-IT FideliTaq PCR Master Mix and stability. A 306 bp fragment of the single-copy numb gene was amplified from 1 ng of human genomic DNA with and without HotStart-IT technology. The primers in this assay were designed with 3 bases of overlap at their 3'-ends to favor primer-dimer formation during reaction set-up at room temperature. Results demonstrate a shift from mainly primer-dimers to the desired product when HotStart-IT is used. Also, the mix is extremely stable as no loss in performance was observed following 10 freeze-thaw cycles (FTs) relative to a mix stored at -20°C.
Fig. 2. Sensitivity of the HotStart-IT FideliTaq PCR Master Mix. A 455 bp fragment of the single-copy numb gene was amplified from the indicated amounts of human genomic DNA. The master mix is extremely sensitive as amplification can be achieved from approxi-mately one human cell.
Fig. 3. Extreme amplicon length using HotStart-IT FideliTaq PCR Master Mix and stability. Two long PCR products of 20.7 kb and 35 kb were amplified from lambda DNA using 1 ng and 10 ng of DNA respectively. A 23 kb fragment of the β-globin gene was ampli-fied from 100 ng of human genomic DNA. A denaturation tempera-ture of 95°C was used for lambda targets and 92°C was used for the human target. Note that the mix is extremely stable as no loss in per-formance was observed following 10 freeze-thaw cycles (FTs) relative to a mix stored at -20°C.
500 bp –
100 bp –
no HotStart
HotStart
– numb, 306 bp
– primer-dimers
500 bp –
100 bp –
3 pg (1 cell)
30 pg
– numb, 455 bp
300 pg
19For bulk or alternate pack sizes, email us at [email protected].
PCR Tools | Hot Start PCR
71195 50 units 250 units 1,000 units 5 x 250 units 5,000 unitsCustoms by request
Source:E. coli strain expressing a clone of Taq DNA Polym-erase from Thermus aquaticus(1-3). The hot start component is a recombinant protein also expressed in E. coli.
Description:HotStart-IT Taq DNA Polymerase uses a novel hot start method developed at Affymetrix called primer sequestration. In general, hot start PCR methods reduce or eliminate non-specific primer-extension products formed at lower temperatures during assembly of PCR reactions. At these less strin-gent annealing temperatures, primers may bind non-specifically, which often leads to unwanted amplification products and primer-dimers. In order to resolve this problem, we have combined Taq DNA Polymerase with a recombinant protein which binds and sequesters primers at lower tempera-tures making them unavailable for use by Taq DNA Polymerase. This primer-sequestration technique effectively blocks DNA synthesis from mis-priming events at lower temperatures (Fig. 1). Following the initial denaturation step during PCR, the protein is inactivated and the primers are free to participate in the amplification reaction. This novel hot start method enhances many complex PCR reactions by increasing both specificity and yield.
Applications:�� High-specificity PCR amplification�� High-sensitivity PCR amplification�� TA-vector cloning�� Amplification prior to in vitro transcription
Advantages:�� Room temperature reaction set-up�� High specificity and sensitivity�� Minimizes amplification of non-specific products and primer-dimers (Fig. 2)
Properties:Activator: Mg2+
Taq DNA Polymerase is a highly processive 5'→3' polymerase and has 5'→3' exonuclease activity. Suitable for TaqMan® assays.
Purity:Free from detectable non-specific nucleases.
Storage buffer:20 mM Tris-HCl, pH 8.5, 1 mM DTT, 0.1 mM EDTA, 200 mM KCI, 50% glycerol, stabilizers.
Unit definition:One unit incorporates 10 nmol of total nucleotides into acid-insoluble material in 30 minutes at 74°C in a total volume of 50 µl.
Concentration:1.25 units/µl
Assay conditions:The reaction mixture (50 µl) contains 25 mM TAPS, pH 9.3 (at 25°C), 50 mM KCl, 2 mM MgCl2, 1 mM 2-mercaptoethanol, 200 µM dNTPs, 250 µg/ml activated salmon sperm DNA, and cloned Taq DNA Polymerase. After incubation at 74°C for 10 minutes, acid-insoluble material is determined.
Polymerase blocking assay:The assay compares the polymerase activity of HotStart-IT Taq DNA Polymerase relative to Taq DNA Polymerase (PN 71160). The reaction mixtures contain 0.625 units of polymerase, 1X PCR Reaction Buffer, 0.2 mM each dNTP, and 2 pmol of overlap-ping, extendable oligonucleotides in a 25 µl reaction volume. Following incubation at 25°C for 4 hours, HotStart-IT Taq DNA Polymerase blocks at least 90% of the activity relative to Taq DNA Polymerase without hot start capability.
Functional test:PCR with HotStart-IT Taq DNA Polymerase shifts production of primer-dimers to a specific target of 306 bp from 1 ng of human genomic DNA relative to Taq DNA Polymerase.
Functionally tested 10X PCR Reaction Buffer (included):100 mM Tris-HCl (pH 8.6), 500 mM KCl, 15 mM MgCl2
Functionally tested MgCl2 (included):25 mM solution
Shipping and storage:Shipped on dry ice. Store at -20°C.
References:1. Innis, M. A., Myambo, K. B., Gelfand, D. H. and
Brow, M. A. (1988) Proc. Natl. Acad. Sci. 85, 9436-9440.
2. Lawyer, F. C., Stoffel, S., Saiki, R. K., Myambo, K., Drummond, R. and Gelfand, D. H. (1989) J. Biol. Chem. 264, 6427-6437.
3. Innis, M. A. and Gelfand, D. H. (1990) PCR Protocols: A Guide to Methods and Applications, Academic Press.
HotStart-IT Taq DNA Polymerase
100 pg DNA 1 ng DNA - - + + M - - + +
– numb, 306 bp– primer- dimers
Fig. 2. Increased specificity of HotStart-IT Taq DNA Polymerase. The single-copy numb gene was amplified from the indicated amounts of human genomic DNA with standard Taq DNA Polymerase (-) or with HotStart-IT Taq DNA Polymerase (+). Primers were designed with a 3 bp overlap at their 3'-ends to favor primer-dimer formation during reaction set-up at room temperature. Results demonstrate a shift from mainly primer-dimers to the desired product when HotStart-IT Taq DNA Polymerase is used.
Fig. 1. HotStart-IT Method: Primer Sequestration
PCR Reaction Preparation without Hot Start
• primers with self-complementary region
5 3
5 3
5 3
553
5 3
• primers hybridize to each other at low temperatures
PCR Reaction Preparation with USB Hot Start Method
5 3
5 3
53
5 3
5 3
53
• polymerases are recruited to hybrid• primer-dimers are produced• PCR Failure
• binding proteins interact with primers• primers are sequestered at low temperatures
• prevents non-specific hybridization and extension
• denaturation step inactivates binding proteins• primers bind to specific targets
• PCR Success
3Polymerase
Polymerase
• primers with self-complementary region
Binding Proteins
20 888-362-2447 | 216-765-5000 | usb.affymetrix.com
71196 25 reactions 100 reactions 500 reactionsCustoms by request
HotStart-IT Taq Master Mix uses a novel hot start method developed at Affymetrix called primer sequestration. In general, hot start PCR methods reduce or eliminate non-specific primer-extension products formed at lower temperatures during assembly of PCR reactions. At these less stringent annealing temperatures, primers may bind non-specifically, which often leads to unwanted amplification products and primer-dimers.
In order to resolve this problem, we have combined Taq DNA Polymerase with a recombinant protein which binds and sequesters primers at lower temperatures making them unavailable for extension by Taq DNA Polymerase. This primer-sequestration technique effectively blocks DNA synthesis from mis-priming events at lower temperatures (Fig. 1). Following the initial denaturation step during PCR, the protein is inactivated and the primers are free to participate in the amplification reaction. This novel hot start method enhances many complex PCR reac-tions by increasing both specificity and yield.
HotStart-IT Taq Master Mix is supplied as a conve-nient 2X pre-mixed formulation containing HotStart-IT Taq DNA Polymerase and Ultrapure nucleotides in a proprietary reaction buffer.
Applications:�� High-specificity PCR amplification�� High-sensitivity PCR amplification�� TA-vector cloning�� Amplification prior to in vitro transcription�� High-throughput PCR
ConvenientThe pre-mixed formulation saves time and reduces potential contamination errors by eliminating several pipetting steps. For a 50 µl reaction, simply add 25 µl of HotStart-IT Taq Master Mix to primers, DNA template, and PCR-Qualified H2O. Reactions can be easily performed in 10 µl, 25 µl, 50 µl or 100 µl volumes.
Room temperature reaction assembly is possible because of the hot start feature.
Novel hot start technologySince the polymerase is not chemically-inactivated, there is no extensive heating step necessary which reduces the chance of damaging precious DNA samples from heat-induced depurination.
Higher specificity and sensitivityHotStart-IT Taq Master Mix minimizes amplifica-tion of non-specific products and primer-dimers (Figs. 1 and 2). It amplifies PCR products with low background and from low-copy targets, essential for demanding genomic and cDNA applications with limited sample material.
StableThe mix withstands repeated freeze-thaw cycles and 4°C storage for extended periods of time with no observed decrease in performance (Fig. 2).
Functional test:PCR with HotStart-IT Taq Master Mix shifts produc-tion of primer-dimers to a specific target of 306 bp from 1 ng of human genomic DNA relative to Taq Master Mix.
HotStart-IT Taq Master Mix Formulation (2X):HotStart-IT Taq Master Mix combines Taq DNA Polymerase with a recombinant hot start protein in a unique buffer formulation. Magnesium and nucleotide concentrations are 3 mM and 0.4 mM each, respectively.
Components:25 reactions 625 µl100 reactions 4 x 625 µl500 reactions 12.5 ml
Shipping and storage:Shipped on dry ice. Store at -20°C. Mix well prior to use.
PCR Tools | Hot Start PCR
HotStart-IT Taq PCR Master Mix (2X)
– numb, 306 bp
– primer- dimers
M - + + + +-20
°CF/T 4°C RT
Fig. 2. Increased specificity and stability of HotStart-IT Taq Master Mix. Specificity: The single-copy numb gene was amplified from 1 ng of human genomic DNA with standard Taq Master Mix (-) or with HotStart-IT Taq Master Mix (+). Primers were designed with a 3 bp overlap at their 3'-ends to favor primer-dimer formation during reaction set-up at room temperature. Results demonstrate a shift from mainly primer-dimers to the desired product when HotStart-IT Taq Master Mix is used. Stability: The Master Mix shows no loss in performance following 15 freeze-thaw cycles (F/T), 4°C storage for one month (4°C), or room temperature storage for one month (RT) compared to a reference mix stored at -20°C.
4 hours at 24 hours at 25°C 25°C - Taq HS - Taq HS
– 50-mer extended product23-mer
template –
Fig. 1. HotStart-IT Taq Master Mix prevents primer exten-sion at room temperature. Two overlapping and extendable oligonucleotides were incubated in a mock PCR reaction at 25°C for either 4 or 24 hours with standard Taq Master Mix (Taq), HotStart-IT Taq Master Mix (HS), or no polymerase (-). Following incubation, reactions were separated on a 15% denaturing gel with urea. Results demonstrate that full-length extension of the HEX-labeled 23-mer to a 50-mer was completely blocked by HotStart-IT Taq Master Mix at room temperature while some extension occurred with the standard Taq Master Mix.
Related products
ExoSAP-IT PCR Product Cleanup78250 20 reactions78200 100 reactions78201 500 reactions78202 2,000 reactions78205 5,000 reactions
PCR Markers, 50 - 2000 bp76710 250 µl
PCR Nucleotide Mix, 10 mM Solution(10 mM each dATP, dCTP, dGTP, dTTP)77212 500 µl
Water, Nuclease-Free71786 10 x 1 ml 100 ml 500 ml 1 L 5 L
21For bulk or alternate pack sizes, email us at [email protected].
71194 400 μgSufficient for 200 x 25 μl reactionsCustoms by request
Source:HotStart-IT Binding Protein is a recombinant protein expressed in E. coli.
Description:HotStart-IT Binding Protein is the active compo-nent in a novel hot start technology developed at Affymetrix called primer sequestration. In general, hot start PCR methods reduce or eliminate non-specific primer-extension products formed at lower temperatures during assembly of PCR reactions. At these less stringent annealing temperatures, primers may bind non-specifically, which often leads to unwanted amplification products and primer-dimers. In order to resolve this problem, we have produced a high-quality DNA binding protein that is especially useful at sequestering primers at lower temperatures making them unavailable for use by a polymerase (Fig. 1). This primer-sequestration technique effectively blocks DNA synthesis from mis-priming events at lower temperatures. Following the initial denaturation step during PCR, the protein is inactivated and the primers are free to participate in the amplification reaction. Unlike other hot start methods, such as antibodies or chemical modifica-tions, the binding protein is compatible with a vari-ety of thermostable polymerases. HotStart-IT Binding Protein has been designed for PCR applications that demand extremely high specificity and sensitivity and is thoroughly tested for purity and performance.
Application:�� Hot start PCR amplification
Properties:HotStart-IT Binding Protein performs well in many standard PCR reaction buffers.
Purity:Free from detectable non-specific nucleases.
Storage buffer:20 mM Tris-HCl (pH 8.5), 200 mM KCl, 0.1 mM EDTA, 1 mM DTT, and 50% glycerol.
Mass definition:In general, one microgram of HotStart-IT Binding Protein sequesters about 5 pmol of primers.
Concentration:≥ 10 mg/ml
Quality control polymerase blocking assay:This assay compares the amount of Taq DNA Polymerase activity with 2 μg HotStart-IT Binding Protein against its activity with no binding protein. The reaction mixtures contain 0.625 units of polymerase, 1X PCR Reaction Buffer (10 mM Tris- HCl [pH 8.6], 50 mM KCl, 1.5 mM MgCl2), 0.2 mM each dNTP, and 2 pmol of overlapping, extendable oligonucleotides in a 25 µl reaction volume. Following incubation at 25°C for 4 hours, HotStart-IT Binding Protein blocks at least 90% of the Taq DNA Polymerase activity.
Functional test:PCR with Taq DNA Polymerase and HotStart-IT Binding Protein shifts production of primer-dimers to a specific target of 306 bp from 1 ng of human genomic DNA relative to Taq DNA Polymerase alone.
Shipping and storage:Shipped on dry ice. Store at -20°C.
PCR Tools | Hot Start PCR
HotStart-IT Binding Protein
Fig. 2. Increased specificity due to HotStart-IT Binding Protein. Single-copy numb and p53 genes were amplified from the indicated amounts of human genomic DNA using standard Taq DNA Polymerase without (-) and with (+) 2 µg of HotStart-IT Binding Protein per reaction. The primers in this assay were designed with 3 bases of overlap at their 3'-ends to favor primer-dimer formation dur-ing reaction set-up at room temperature. Results demonstrate a shift from mainly primer-dimers to the desired products when the binding protein was included.
Fig. 1. HotStart-IT Method: Primer Sequestration
PCR Reaction Preparation without Hot Start
• primers with self-complementary region
5 3
5 3
5 3
553
5 3
• primers hybridize to each other at low temperatures
PCR Reaction Preparation with USB Hot Start Method
5 3
5 3
53
5 3
5 3
53
• polymerases are recruited to hybrid• primer-dimers are produced• PCR Failure
• binding proteins interact with primers• primers are sequestered at low temperatures
• prevents non-specific hybridization and extension
• denaturation step inactivates binding proteins• primers bind to specific targets
• PCR Success
3Polymerase
Polymerase
• primers with self-complementary region
Binding Proteins
Advantages of HotStart-IT:�� Room temperature reaction set-up�� High specificity and sensitivity�� Minimizes amplification of non-specific products and primer-dimers (Fig. 2).�� Ideal for complex templates and multiplex reactions�� Unlike chemically-modified Taq, no extensive heating step is necessary which may damage precious samples.�� Technology is portable to a polymerase of choice.
– numb, 306 bp– primer-dimers
– p53, 1142 bp
– primer-dimers
- - + + M - - + +
- - + + M - - + +
22 888-362-2447 | 216-765-5000 | usb.affymetrix.com
PCR Tools | Multiplex PCR
71199 25 reactions 100 reactions
MagniTaq Multiplex PCR Master Mix is a single-tube, 2X multiplex PCR master mix with validated perfor-mance for simultaneous amplification of at least 20 targets up to 1 kb in length of difficult templates with minimal optimization. MagniTaq Multiplex PCR Master Mix (2X) contains hot start MagniTaq DNA Polymerase, dNTPs, and MgCl2 in a proprietary buffer formulation designed to enable multiplex PCR without lengthy optimization procedures.
Features:�� Advanced buffer formulation that includes dNTPs and MgCl2, ensuring efficient, rapid, multiplex PCR without lengthy optimization procedures�� Validated performance for simultaneous amplification of at least 20 targets up to 1 kb in length�� Quality results on difficult sample types; superior data even from FFPE samples or for single-copy genes�� Single-tube, pre-mixed solution. Simply add template, primers, and water
Applications:�� End point PCR of multiple targets simultaneously�� Minimizes effect of off-target priming events
Reliable and efficient multiplexing of over 20 plexesIn order to enhance the specificity and sensitivity of multiplexing, the mix uses MagniTaq DNA Polymerase, a chemically modified, hot start polymerase that is completely inactive at ambient temperatures. MagniTaq DNA Polymerase minimizes the effect of off-target priming events, such as primer-dimers, which often occur at lower tempera-tures during reaction set-up and prior to the initial denaturation step.
In addition, MagniTaq Multiplex PCR Master Mix efficiently amplifies greater than 20 products simul-taneously over a broad range of template amounts and primer concentrations.
Data generated shows 22-plex amplification from single-copy genes in the Notch signaling pathway. Even at template amounts ranging from 1–100 ng of human gDNA, amplification of 22 targets is observed (Fig. 1). The reaction was done without optimization in a single tube of pre-mixed solution of a proprietary buffer that includes dNTPs and MgCl2.
Varying primer concentrations can affect the ef-ficiency and amplification effectiveness. Typically, lower primer concentrations favor longer products, while higher primer concentrations favor shorter products. MagniTaq Multiplex PCR Master Mix is designed to work over a range of primer concentra-tions (Fig. 2).
Primer design�� The length of primers should be 20-30 bases.�� Aim for a Tm equal to or greater than 60°C; primer sets should be designed to have similar melting temperatures if possible.�� Optimal GC content should be 40-65%.�� Avoid designing primers with 3 or more G or C residues sequentially near the 3’ end, as well as primers that can form hairpins or that have significant complementary sequence to one another.�� Typically, lower primer amounts tend to favor longer targets, while higher primer amounts tend to favor shorter targets.
MagniTaq™ Multiplex PCR Master Mix (2X)
Fig. 1. 22-plex amplification of low amounts of gDNA. 22-plex amplification of gDNA targets with MagniTaq Multiplex PCR Master Mix. 1, 10, 100, and 1000 ng human genomic DNA input amounts (1 ng is about 100 cells) are added to the 22-plex notch-pathway single-copy gene primer set. Primer concentration is 0.2 µM and samples were loaded with SYBR® Green dye and run on an agarose gel.
1 10 100 1000 ng template DNA
1000900800700600
500
400
300
200
100
1252 1252
142 142
1500
1000
750
500
300
150
50 Fig. 2. 22-plex of a wide range of primer concentrations. MagniTaq Multiplex PCR Master Mix is effective over a wide range of primer concentrations. 100 ng human genomic DNA input amount is added to the 22-plex notch-pathway, single-copy gene primer set. Duplicate reactions were run using 0.1, 0.2 or 0.4 µM primer concentrations. Samples were loaded with SYBR Green dye and run on an agarose gel.
µM primers: 0.1 0.2 0.4
1000900800700600
500
400
300
200
100
1252
142
1500
1000
750
500
300
150
50
23For bulk or alternate pack sizes, email us at [email protected].
PCR Tools | Multiplex PCR
Fig. 4. Comparison testing of 22-plex FFPE samples. 22-plex FFPE sample amplification competitor comparison data of normal vs tumor sample types. 100 ng human genomic DNA input from 2008 breast FFPE samples and 2002 lung FFPE samples, normal or tumor, are added to the 22-plex notch pathway primer set. 0.1 µM primer sets are used. Comparison of MagniTaq Multiplex PCR Master Mix (MT) to competitor Q and QP were run. Samples are loaded with SYBR Green and run on 2% TAE-agarose gel.
Fig. 3. Superior multiplexing of FFPE samples. 20-plex FFPE sample amplification competitor comparison data of normal vs. tumor sample types. 25 ng human genomic DNA input from 2008 breast FFPE samples and 2002 lung FFPE samples, normal or tumor, are added to the 20-plex notch pathway primer set. 0.2 µM primer sets are used. Comparison of MagniTaq Multiplex PCR Master Mix (MT) to competitor Q and QP were run. Samples are loaded with SYBR Green and run on 2.25% TAE-agarose gel.
Breast—2,008 Lung—2,002
50 ng FFPE gDNANormal (N) or Tumor (T)
MT Q QP MT Q QP N T N T N T N T N T N T
1000900800700600500400300
200
100
1500
1000
750
500
300
150
50
1252
142
25 ng FFPE gDNANormal (N) or Tumor (T)
MT Q QP MT Q QP N T N T N T N T N T N T
MagniTaq™ Multiplex PCR Master Mix (2X) continued...
Superior performance on difficult targets with minimal optimizationMagniTaq Multiplex PCR Master Mix offers quality results on difficult to amplify sample types. Genomic DNA isolated from FFPE tissue samples is often degraded, and therefore, difficult to amplify using standard PCR reagents. Multiplex PCR with these sample types calls for an exceptional amount of optimization to produce consistent amplification of all sought after targets.
The uniform product yield obtained when using MagniTaq Multiplex PCR Master Mix allows for more products to be amplified, especially when compared to mixes from other suppliers (Figs. 3 and 4). The mixes from other suppliers show a much more pronounced decrease in the amplification of the high molecular weight products compared to MagniTaq Multiplex PCR Master Mix (Figs. 3 and 4). MagniTaq Multiplex PCR Master Mix is, therefore, more suit-able for PCR amplification of multiple primer sets from degraded FFPE gDNA template, particularly for older tissue samples.
Components:25 reactions—for 25 x 50 μl multiplex PCR
reactions100 reactions—for 100 x 50 μl multiplex PCR
reactions
Shipping and storage:Shipped on dry ice. For long-term storage, store at -20°C. When using, the mix may be stored at 4°C for up to 2 months.
24 888-362-2447 | 216-765-5000 | usb.affymetrix.com
PCR Tools | Multiplex PCR
MP1100 kitPCR Primers*MP11XX *Use digits from the PCR Primers product number. See chart below for full listing of PCR Primers.
�� Quickly and easily detect the methylation status of your promoter of interest�� No hassle of sodium bisulfate modification�� Choose from our list of 40 PCR primers OR use your own PCR primers to detect methylation of promoters specific for your research.
Use the Promoter Methylation PCR Kit to quickly and easily detect the methylation status of your promoter of interest, without the hassle of chemical modifica-tion with sodium bisulfate.
Standard molecular biology techniques do not preserve methylation of the genomic DNA, causing a hindrance in the appreciation of methylation as an important epigenetic event in cancer progression. Affymetrix has developed a PCR method for analyz-ing methylation status of a specific promoter of interest. The method is based on MeCP2 bind-ing, which differentiates those promoters with methylated groups from unmethylated promoters. Compared to sodium bisulphate-based conversion of methylated bases in conjunction with PCR amplifica-tion, this method is very simple and straightforward.
The flowchart below details how this procedure works:Isolated genomic DNA is digested with Mse I, and the resulting DNA fragments are incubated with the
methylation binding protein MeCP2 (a.k.a. MBP). The methylated DNA fragments are isolated with a spin column and then amplified with promoter specific primers. Agarose gel electrophoresis is used to visualize the PCR products. The presence of a band on the gel indicates that a specific promoter is methylated in your genomic DNA sample.
Mse I and PCR enzymes are not included in this kit. We recommend HotStart-IT FideliTaq DNA Polymerase, PN 71155 and MagniTaq Multiplex PCR Master Mix, PN 71199, for PCR (see below).
Promoter Methylation PCR Kit
PCR primers:Gene Name Forward Reverse Approx. PCR Prod. No. Primer Position Primer Position Product Length14-3-3 1 180 bp MP1101 “ 180 “ MP1102
14-3-3 300 600 bp MP1103 “ 900 “ MP1104
ABL1 2 390 bp MP1105 “ 400 “ MP1106
BAGE 1 230 bp MP1107 “ 230 “ MP1108
BAGE 260 490 bp MP1109 “ 650 “ MP1110
BAGE 670 230 bp MP1111 “ 900 “ MP1112
BRCA1 50 200 bp MP1113 “ 250 “ MP1114
BRCA1 550 130 bp MP1115 “ 680 “ MP1116
Calcitonin CGPR 1 300 bp MP1117 “ 450 “ MP1118
Calcitonin CGPR 670 200 bp MP1119 “ 900 “ MP1120
CD14 540 400 bp MP1121 “ 1000 “ MP1122
CDKN2A 900 200 bp MP1123 “ 1100 “ MP1124
Gene Name Forward Reverse Approx. PCR Prod. No. Primer Position Primer Position Product LengthCDKN2A 270 240 bp MP1125 “ 510 “ MP1126
COX2 280 320 bp MP1127 “ 601 “ MP1128
CyclinD2 580 170 bp MP1129 “ 750 “ MP1130
CyclinD2 440 130 bp MP1131 “ 570 “ MP1132
DAPK 50 200 bp MP1133 “ 250 “ MP1134
DAPK 400 500 bp MP1135 “ 900 “ MP1136
glut4 190 200 bp MP1137 “ 390 “ MP1138
H-ras 40 400 bp MP1139 “ 460 “ MP1140
H-ras 500 300 bp MP1141 “ 840 “ MP1142
INFg 500 100 bp MP1143“ 600 “ MP1144
GAGE1 100 250 bp MP1145 “ 390 “ MP1146
GAGE1 400 400 bp MP1147 “ 800 “ MP1148
Fig. 1. Methylation PCR verification of array results. Results obtained with the Promoter Methylation Array can be verified with the Promoter Methylation PCR Kit. Genomic DNA from MCF7 and HeLa cells was analyzed with the Promoter Methylation Array (top). The promoters (indicated as boxes) were verified by individual meth-ylation PCR. Left band: MCF7. Right band: HeLa. (bottom).
Related products
HotStart-IT FideliTaq DNA Polymerase71155 50 units 250 units 1,000 units 5,000 units
MagniTaq Multiplex PCR Master Mix (2X)71199 25 reactions 100 reactions
25For bulk or alternate pack sizes, email us at [email protected].
PCR Tools | Multiplex PCR
Promoter Methylation PCR Kit continued...
71155 50 units 250 units 1,000 units 5,000 unitsCustoms by request
Source:All proteins are recombinant versions expressed in E. coli.
Description:HotStart-IT FideliTaq DNA Polymerase combines a novel hot start method developed at Affymetrix with the long and accurate amplification properties of FideliTaq DNA Polymerase. The new hot start method, called primer sequestration, uses a binding protein to reduce or eliminate non-specific primer-extension products which may be generated at lower temperatures during assembly of PCR reactions. Following the initial denaturation step during PCR, the binding protein is inactivated and the primers are free to participate in the amplification reaction. This novel method enhances many complex PCR reactions by increasing specificity and yield, as well as sensitivity.
FideliTaq DNA Polymerase combines Taq DNA Polym-erase with a high-fidelity, proofreading polymerase. This enzyme blend has the 5'→3' exonuclease activity of Taq DNA Polymerase as well as the 3'→5' exonuclease activity of the proofreading enzyme. FideliTaq DNA Polymerase increases fidelity up to 6-fold over Taq DNA Polymerase alone and allows for amplification of longer products(1-4). The enzyme blend generates PCR products whose ends are compatible with either blunt-end or TA cloning procedures(5), with A-tailed ends favored over blunt ends in an approximately 3 to 1 ratio.
FideliTaq DNA Polymerase can be used in conjunc-tion with the Promoter Methylation PCR Kit for gene amplification prior to methylation detection.
Applications:�� High-specificity and high-sensitivity PCR amplification�� Extremely long PCR amplification�� PCR-mediated cloning
Advantages:�� Room temperature reaction set-up.�� Minimizes amplification of non-specific products and primer-dimers.�� High specificity and sensitivity �� Generates long PCR amplicons�� Up to 6-fold higher fidelity than Taq polymerase.
Properties:Activator: Mg2+
Purity:Free from detectable non-specific nucleases.
Storage buffer:20 mM Tris-HCl (pH 8.5), 1 mM DTT, 0.1 mM EDTA, 200 mM KCI, 50% glycerol, and stabilizers.
Unit definition:One unit incorporates 10 nmol of total nucleotides into acid-insoluble material in 30 minutes at 74°C in a total volume of 50 µl(6-7).
Concentration:2.5 units/µl
Assay conditions:The reaction mixture contains 25 mM TAPS, pH 9.3 (at 25°C), 50 mM KCl, 2 mM MgCl2, 1 mM 2-mercaptoethanol, 200 µM dNTPs, 250 µg/ml activated salmon sperm DNA, and Taq DNA Polymerase. Following incubation at 74°C for 10 minutes, acid-insoluble material is determined.
Functional tests:PCR with HotStart-IT FideliTaq DNA Polymerase shifts production of primer-dimers to a specific target of 306 bp from 1 ng of human genomic DNA rela-tive to FideliTaq DNA Polymerase.
PCR with HotStart-IT FideliTaq DNA Polymerase generates a 20.7 kb product.
Functionally tested 10X PCR Reaction Buffer (included): 100 mM Tris-HCl (pH 8.6), 500 mM KCl, 15 mM MgCl2Functionally tested MgCl2 (included):25 mM solution
Shipping and storage:Shipped on dry ice. Store at -20°C.
References:1. Barnes, W. M. (1994) Proc. Natl. Acad. Sci. USA
91, 2216-2220. 2. Cheng, S., Fockler, C., Barnes, W. M., and
Higuchi, R. (1994) Proc. Natl. Acad. Sci. USA 91, 5695-5699.
3. Barnes, W. M. (1992) Gene 112, 29-35. 4. Cline, J., Braman, J. C., and Hogrefe, H. H. (1996)
Nucl. Acids Res. 24, 3546-3551. 5. Magnuson, V. L., Ally, D. S., Nylund, S. J.,
Karanjawala, Z. E., Rayman, J. B., Knapp, J. I., Lowe, A. L., Ghosh, S., and Collins, F. S. (1996) BioTechniques 4, 700-709.
6. Innis, M. A., Myambo, K. B., Gelfand, D. H. and Brow, M. A. (1988) Proc. Natl. Acad. Sci. 85, 9436-9440.
7. Lawyer, F. C., Stoffel, S., Saiki, R. K., Myambo, K., Drummond, R. and Gelfand, D. H. (1989) J. Biol. Chem. 264, 6427-6437.
HotStart-IT FideliTaq DNA Polymerase
PCR primers continued:Gene Name Forward Reverse Approx. PCR Prod. No. Primer Position Primer Position Product LengthHOXA2 1 300 bp MP1149 “ 300 “ MP1150
IL4 450 230 bp MP1151 “ 680 “ MP1152
IRF7 350 450 bp MP1153 “ 850 “ MP1154
NIS 1 350 bp MP1155 “ 400 “ MP1156
NIS 500 300 bp MP1157 “ 800 “ MP1158
NME2 80 170 bp MP1159 “ 250 “ MP1160
NME2 280 500 bp MP1161 “ 800 “ MP1162
PDGFB 1 500 bp MP1163 “ 678 “ MP1164
Gene Name Forward Reverse Approx. PCR Prod. No. Primer Position Primer Position Product LengthPOMC 50 490 bp MP1165 “ 540 “ MP1166
NES 150 290 bp MP1167 “ 410 “ MP1168
NES 450 350 bp MP1169 “ 800 “ MP1170
PRA2 400 250 bp MP1171 “ 650 “ MP1172
TFF1 510 160 bp MP1173 “ 750 “ MP1174
VHL 80 290 bp MP1175 “ 380 “ MP1176
Survivin 1 300 bp MP1177 “ 300 “ MP1178
CASP8 180 360 bp MP1179 “ 640 “ MP1180
26 888-362-2447 | 216-765-5000 | usb.affymetrix.com
PCR Tools | Real-Time qPCR
71170 50 units 250 units 1,000 units 5,000 units
�� Chemically-modified Taq polymerase for hot start PCR�� Minimizes the amplification of non-specific products and primer dimers.�� Exceptional performance, offering high specificity and sensitivity�� Room temperature reaction set-up
VeriQuest Taq DNA Polymerase is a highly purified, hot start Taq polymerase that has no polymerase activity prior to the initial heat activation step, ensur-ing there is no amplification of non-specific primers, such as primer-dimers. This feature allows the con-venience of reaction assembly at room temperature as well as higher specificity and sensitivity. The initial incubation step of 95°C for 10 minutes before PCR cycling removes the blocking chemical moiety, result-ing in activation of the polymerase.
Properties:VeriQuest Taq DNA Polymerase is a chemically-modified, full length Taq DNA polymerase for hot start.
Purity:Free from detectable non-specific nucleases.
Storage buffer:20 mM Tris-HCl, pH 8.5, 100 mM KCl, 1 mM DTT, 0.1 mM EDTA, 50% glycerol and stabilizers.
Unit definition: One unit of enzyme is defined as the amount that catalyzes the incorporation of 10 nmol of total nucleotide into acid-insoluble form in 30 minutes at 74°C.
Assay conditions: The reaction mixture contains 25 mM TAPS, pH 9.3 (at 25°C), 50 mM KCl, 2 mM MgCl2, 1 mM β-ME, 200 µM each dATP, dGTP, dTTP, 100 µM [α-32P]-dCTP (0.05 to 0.1 Ci/mmole), 250 µg/ml activated salmon
sperm DNA, and VeriQuest Taq DNA Polymerase. After incubation at 74°C for 10 minutes, acid insoluble material is determined (50 µl reaction volume).
Functional test: Functionally tested for PCR according to the stan-dard Affymetrix PCR protocol. This protocol is based upon the amplification of a 500 bp lambda DNA.
Concentration: 5 units/µl
Functionally tested VeriQuest Taq 10X PCR Reaction Buffer (included):150 mM Tris-HCl, pH 8.0, 500 mM KCl, 15 mM MgCl2Functionally tested MgCl2 (included):25 mM MgCl2 solution
Storage:Shipped on dry ice. Store at -20°C.
VeriQuest Taq DNA Polymerase
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27For bulk or alternate pack sizes, email us at [email protected].
75600 (50 µl reaction volume) 40 reactions (1 ml) 200 reactions (5 ml) 400 reactions (2 x 5 ml) 1,000 reactions (5 x 5 ml) 2,000 reactions (10 x 5 ml)
VeriQuest SYBR Green qPCR Master Mix is supplied as a 2X pre-mixed formulation containing both ROX and UDG in an optimized buffer for quality results in real-time quantitative PCR assays. The proprietary re-action buffer and the hot start polymerase enhance SYBR Green-based qPCR amplification by reducing primer-dimer formation and non-template priming, increasing specificity and sensitivity. The master mix contains a chemically-modified Taq DNA Polymerase, MgCl2, ultrapure nucleotides with an optimized dUTP:dTTP ratio, Uracil-DNA Glycosylase (UDG or UNG), SYBR Green I, and ROX Passive Reference Dye in a proprietary reaction buffer.
Features:�� One-tube master mix with ROX Reference Dye included�� Same protocols and same primers for standard mode amplification�� Consistency over a broad dynamic range: 7 orders of magnitude linear detection range with a correlation coefficient ≅ 1 (see Fig. 1) �� Exceptional performance with challenging GC-rich regions (see Fig. 3)�� Detect less than 4 copies of target from genomic DNA and differentiate less than a two-fold difference.�� Contains dUTP and Uracil-DNA Glycosylase (UDG or UNG) for carryover contamination prevention.�� Hot start PCR for room temperature reaction setup�� Stable at room temperature for 72 hours in a preassembled reaction
Reproducible, consistent results while main-taining precision and efficiencyThe convenience of a master mix formulation without sacrificing quality. VeriQuest SYBR Green qPCR Master Mix provides consistency over a broad dynamic template concentration range allowing 7 orders of magnitude linear detection (Fig. 1). High precision in target quantification and discrimination of a 1.33 to 10-fold dilution series with VeriQuest SYBR Green qPCR Master Mix ensure accurate results.
Enhanced amplification sensitivity and accuracy of low copy number detectionReliable detection from as little as 4 copies of a single copy gene and differentiation from less than a two-fold difference (Fig. 2).
Exceptional performance with challenging templatesEven in high GC and AT rich regions, VeriQuest SYBR Green qPCR Master Mix offers exceptional performance and high specificity so you have the confidence to verify your gene expression results (Figs. 3 and 4).
Ready-to-use, single tube replacementThe one tube master mix contains all necessary com-ponents: chemically-modified Taq DNA polymerase, MgCl2, ultrapure nucleotides with an optimized dUTP:dTTP ratio, Uracil-DNA Glycosylase (UDG or UNG), SYBR Green I, and ROX as the passive reference dye in a proprietary reaction buffer. Simply add your DNA template, primers, and water, and you can begin your qPCR reaction. For use in your new or existing protocols, on any leading PCR platform, providing easy transition from other master mixes.
Passive Reference Dye and UDG includedROX Passive Reference Dye is an inert dye, whose fluorescence does not change during the reaction. This signal normalization is necessary to correct for well-to-well differences that may occur due to artifacts such as pipetting errors or sample evapora-tion. Uracil-DNA Glycosylase (UDG or UNG) and dUTP offer an option for carry-over contamination prevention from previous PCR amplifications. All are at optimized levels so there is no need to adjust concentrations.
Highly stable and easy to work with VeriQuest SYBR Green qPCR Master Mix is stable at room temperature for 72 hours in a pre-assembled reaction and can be stored at 4°C for convenient handling (Figs. 5 and 6); no lost time waiting for your master mix to thaw. Testing of 10 freeze-thaw cycles showed no loss in master mix performance. Ideal for high-throughput handling.
Components:40 reactions: 1 ml200 reactions: 5 ml400 reactions: 2 x 5 ml1,000 reactions: 5 x 5 ml2,000 reactions: 10 x 5 ml
References:1. VanGuilder, H. D., Vrana, K. E., and Freeman, W. M.
(2008) BioTechniques 44 (5), 619-626.2. Longo, M. C., Berninger, M. S., and Hartley, J. L.
(1990) Gene 93, 125-128.3. Tewhey, R. et al. (2009) Nature Biotechnology 27,
1025-1031.
VeriQuest SYBR Green qPCR Master Mix (2X)
PCR Tools | Real-Time qPCR
Fig. 2. High precision in target quantification with VeriQuest SYBR Green qPCR Master Mix. Amplification plot from real-time PCR for a 1.33 to 10-fold dilution series of 10 ng to 1 ng of cDNAs reverse-transcribed from HeLa total RNA.
Fig. 1. Linear detection range of VeriQuest SYBR Green qPCR Master Mix. Real-time amplification plot from a 10-fold dilu-tion series of a GAPDH synthetic target with starting amounts of 109 copies amplified in four replicate reactions using the ABI StepOne™ Real-Time PCR System.
(Continued on next page)
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PCR Tools | Real-Time qPCR
VeriQuest SYBR Green qPCR Master Mix continued...
Fig. 4. Melt curve from a 71.2% GC target amplicon. Human male genomic DNA was used.
Fig. 3. Amplification of GC-rich target with VeriQuest SYBR Green qPCR Master Mix. Standard curves from a 10-fold dilution series of 100 ng to 10 pg human male genomic DNA amplified in four replicate reactions with VeriQuest SYBR Green qPCR Master Mix, BioRad iTaq™ Fast SYBR Green Supermix with ROX and ABI Power® SYBR Green PCR Master Mix using the ABI 7500 PCR System for detection of a 71.2% GC-target amplicon (PROC, NT_022135.16)(3).
USB VeriQuest
BioRad
ABI
-2.5 -1.5 -0. 5 0.5 1.5 2.5
log gDNA input
44
39
29
34
Ct
19
24
PROC, 71.2% GCMix USB VQ BioRad ABIR2 0.999 0.944 0.997Slope -3.36 -1.75 -3.28EPCR 98.2% 272.6% 101.7%
Fig. 5. VeriQuest SYBR Green qPCR Master Mix stability for reliable performance of high-throughput handling. Pre-assembled PCR reaction stability after 48 and 72 hour incubations at room temperature from quantification of GADPH cDNAs.
Fig. 6. Inset defining Ct levels of 1 ng and 10 pg of cDNAs reverse transcribed from HeLa total RNA. GADPH was detect-ed from preassembled PCR reactions incubated at room temperature for 48 and 72 hours.
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29For bulk or alternate pack sizes, email us at [email protected].
75665 (50 µl reaction volume) 40 reactions (1 ml) 200 reactions (5 ml) 400 reactions (2 x 5 ml) 1,000 reactions (5 x 5 ml) 2,000 reactions (10 x 5 ml)
VeriQuest SYBR Green qPCR Master Mix with Fluo-rescein is supplied as a 2X pre-mixed formulation containing both fluorescein and UDG in an opti-mized buffer for quality results in real-time quantita-tive PCR assays. The proprietary reaction buffer and the hot start polymerase enhance SYBR Green-based qPCR amplification by reducing primer-dimer forma-tion and non-template priming, increasing specificity and sensitivity.
Features:�� One-tube master mix with fluorescein added in optimized levels for use in all thermo cyclers that utilize it for signal normalization�� Enhanced amplification sensitivity and accuracy of low copy number detection�� Reproducible, consistent results while maintaining precision and efficiency�� Exceptional performance with challenging templates�� Ready-to-use, single-tube replacement with UDG included�� Highly stable and easy to work with
Enhanced sensitivity on amplification for low copy number detectionReliable detection from as little as four copies of a single copy gene and differentiation from less than a two-fold difference.
Reproducible, consistent results while main-taining precision and efficiencyThe convenience of a master mix formulation with-out sacrificing quality. VeriQuest SYBR Green qPCR Master Mix with Fluorescein provides consistency over a broad dynamic template concentration range allowing 7 orders of magnitude linear detec-tion. High precision in target quantification and discrimination of a 1.33 to 10-fold dilution series with VeriQuest SYBR Green qPCR Master Mix with Fluorescein ensures accurate results.
Exceptional performance with challenging templatesEven in high GC and AT rich regions, VeriQuest SYBR Green qPCR Master Mix with Fluorescein offers excep-tional performance and high specificity so you have the confidence to verify your gene expression results.
Ready-to-use, single-tube replacementThe one-tube master mix contains all necessary components: chemically-modified Taq DNA polymerase, MgCl2, ultrapure nucleotides with an optimized dUTP:dTTP ratio, Uracil-DNA Glycosylase (UDG or UNG), SYBR Green I, and Fluorescein, as the passive reference dye, in a proprietary reaction buffer. Simply add your DNA template, primers, and water and you can begin your qPCR reaction. For use with instruments requiring fluorescein as a passive reference dye, i.e., Bio-Rad iCycler iQ®/ iQ5, Bio-Rad iCycler MyiQ™.
Passive reference dye and UDG includedFluorescein Passive Reference Dye is an inert dye whose fluorescence does not change during the reaction. This signal normalization is necessary to correct for well-to-well differences that may occur due to artifacts such as pipetting errors or sample evaporation.
Uracil-DNA Glycosylase (UDG or UNG) and dUTP offer an option for carry-over contamination prevention from previous PCR amplifications. All are at optimized levels so there is no need to adjust concentrations.
Highly stable and easy to work withVeriQuest SYBR Green qPCR Master Mix with Fluorescein is stable at room temperature for 72 hours in a preassembled reaction. This mix can be stored at 4°C for convenient handling, with no time lost waiting for your master mix to thaw. Testing of 10 freeze-thaw cycles showed no loss in master mix performance. This master mix is ideal for high-throughput handling.
Components:40 reactions: 1 ml200 reactions: 5 ml400 reactions: 2 x 5 ml1,000 reactions: 5 x 5 ml2,000 reactions: 10 x 5 ml
Storage:Shipped on dry ice. Store at -20°C for long-term storage. Store at 4-8°C for short-term storage (≤3 months).
VeriQuest SYBR Green qPCR Master Mix with Fluorescein (2X)
PCR Tools | Real-Time qPCR
Amplification plots and standard curves from real-time PCR for dilution series of a synthetic GAPDH target with starting amounts of 108 copies amplified in triplicate reactions using the Bio-Rad MyiQ™ Single Color Real-Time PCR Detection System.
Linear detection range of VeriQuest SYBR Green qPCR Master Mix with Fluorescein
30 888-362-2447 | 216-765-5000 | usb.affymetrix.com
75690 (20 µl reaction volume) 100 reactions (1 ml) 500 reactions (5 ml) 1,000 reactions (2 x 5 ml) 2,500 reactions (5 x 5 ml) 5,000 reactions (10 x 5 ml)
Features:�� Uses fast mode thermal cycling conditions for results in one-third of the time when compared to standard run protocols.�� One-tube master mix with ROX Reference Dye included – just add template, primers, and water �� Exceptional performance with challenging GC-rich regions�� Contains dUTP and Uracil-DNA Glycosylase (UDG or UNG) for carryover contamination prevention.�� Hot start PCR for room temperature reaction setup�� Sensitivity and precision with limited targets
VeriQuest Fast SYBR Green qPCR Master Mix is op-timized for SYBR Green detection on all instruments that utilize ROX as a passive reference dye and can be run using Fast mode cycling protocols. The 2X ready-to-use master mix contains hot start VeriQuest Taq DNA Polymerase, MgCl2, ultrapure nucleotides with an optimized dUTP:dTTP ratio, Uracil-DNA Glycosylase (UDG or UNG), SYBR Green, and ROX Passive Reference Dye in a proprietary reaction buf-fer. The hot start Taq polymerase has no polymerase activity prior to the initial heat activation step, which allows reaction assembly at room temperature, as well as higher specificity and sensitivity. Since the mix contains dUTP and UDG, carryover contamination
prevention can be performed prior to amplification. VeriQuest Fast SYBR Green qPCR Master Mix offers the same sensitivity and specificity found with our standard mode mixes, producing results in a fraction of the time.
VeriQuest Fast SYBR Green qPCR Master Mix shows exceptional efficiency and high specificity on challenging templates such as high GC- and AT-rich regions. The optimized formulation ensures no sacrifice in quality for increased speed. Performance comparisons highlight the sensitivity and reliable performance of this platform-independent solution. The sensitivity of the master mix allows for discrimi-nation from a 1.33-fold difference in gene target amount detected. In Fig. 2, a 1.33 to 10-fold dilution series of 10 ng to 1 ng of cDNA reverse-transcribed from HeLa total RNA were amplified with an ef-ficiency of >99%.
VeriQuest Fast SYBR Green qPCR Master Mix is highly stable and easy to work with. It remains stable at room temperature for 72 hours in a pre-assembled reaction and can be stored at 4°C for convenient handling. The mix also allows for room temperature reaction set-up. The speed and stability make VeriQuest Fast SYBR Green qPCR Master Mix ideal for high-throughput handling.
Components:100 reactions 1 ml500 reactions 5 ml1,000 reactions 2 x 5 ml2,500 reactions 5 x 5 ml5,000 reactions 10 x 5 ml
Storage conditionsStore at -20°C for long-term storage. Store at 4-8°C for short-term storage (≤ 3 months).
PCR Tools | Real-Time qPCR
VeriQuest Fast SYBR Green qPCR Master Mix (2X)
Fig.1. Linear detection range of VeriQuest Fast SYBR Green qPCR Master Mix. Amplification plot and standard curve from real-time PCR for a dilution series of a synthetic target with starting amounts of 109 copies amplified in four replicate reactions using the ABI 7500 Real-Time PCR System and GAPDH primers in fast mode with 5 minutes activation at 95°C. The amplification process was lin-ear over eight orders of magnitude.
Fig. 2. High sensitivity and precision in limited target quantification. Amplification plot (left) and standard curve (right) from real-time PCR for a 1.33 to 10-fold dilution series of 10 ng to 1 ng of cDNAs reverse-transcribed from HeLa total RNA.
GAPDH Kras
10 43 2 1 ng RNA
10 4 3 2 1 ng RNA
GAPDH2y = -3.3773x + 22.224
EPCR = 97.7%R² = 0.9955
Krasy = -3.2461x + 30.591
EPCR = 103.3%R² = 0.9993
151719212325272931
-0.2 0 0.2 0.4 0.6 0.8 1 1.2
Ct
log input
GAPDH2 Kras
Fig.1. Linear detection range of VeriQuest Fast SYBR Green qPCR Master Mix. Amplification plot and standard curve from real-time PCR for a dilution series of a synthetic target with starting amounts of 109 copies amplified in four replicate reactions using the ABI 7500 Real-Time PCR System and GAPDH primers in fast mode with 5 minutes activation at 95°C. The amplification process was lin-ear over eight orders of magnitude.
15
20
25
30
35
-4.0 -2.0 0.0 2.0
Ct
log ng input
B2M
ABI Fast SYBR
Bio-Rad iTaq Fast
VeriQuest Fast SYBR
VeriQuest Fast SYBRy = -3.1802 + 23.205EPCR = 106.3%R2 = 0.9999
Bio-Rad iTaq Fast SYBRy = -3.3764 + 22.908EPCR = 97.8%R2 = 0.9991
ABI Fast SYBRy = -3.1984 + 23.755EPCR = 105.4%R2 = 0.9995
Fig. 3. Exceptional results with performance comparison. Standard curves from real-time PCR for a 10-fold dilution series of 10 ng to 1 pg cDNA reverse-transcribed from HeLa total RNA amplified in duplicate reactions with VeriQuest Fast SYBR Green qPCR Master Mix, Bio-Rad iTaq Fast SYBR Green Supermix with ROX™, and ABI Fast SYBR Green Master Mix.
VeriQuest Fast SYBRy = -3.1802 + 23.205EPCR = 106.3%R2 = 0.9999
Bio-Rad iTaq Fast SYBRy = -3.3764 + 22.908EPCR = 97.8%R2 = 0.9991
ABI Fast SYBRy = -3.1984 + 23.755EPCR = 105.4%R2 = 0.9995
Fig. 3. Exceptional results with performance comparison. Standard curves from real-time PCR for a 10-fold dilution series of 10 ng to 1 pg cDNA reverse-transcribed from HeLa total RNA amplified in duplicate reactions with VeriQuest Fast SYBR Green qPCR Master Mix, Bio-Rad iTaq Fast SYBR Green Supermix with ROX™, and ABI Fast SYBR Green Master Mix.
31For bulk or alternate pack sizes, email us at [email protected].
VeriQuest Fast SYBR Green qPCR Master Mix with Fluorescein (2X)
75675 (20 µl reaction volume) 100 reactions (1 ml) 500 reactions (5 ml) 1,000 reactions (2 x 5 ml) 2,500 reactions (5 x 5 ml) 5,000 reactions (10 x 5 ml)
Features:�� Uses fast mode thermal cycling conditions for results in one-third of the time when compared to standard run protocols.�� One-tube master mix with fluorescein passive reference dye included – for use in BioRad thermocyclers �� Exceptional performance with challenging GC-rich regions�� Contains dUTP and Uracil-DNA Glycosylase (UDG or UNG) for carryover contamination prevention.�� Hot start PCR for room temperature reaction setup�� Sensitivity and precision with limited targets
VeriQuest Fast SYBR Green qPCR Master Mix with Fluorescein is optimized for SYBR Green detec-tion on BioRad instruments that utilize fluorescein as a passive reference dye and can be run using Fast mode cycling protocols. The 2X ready-to-use master mix contains hot start VeriQuest Taq DNA Polymerase, MgCl2, ultrapure nucleotides with an optimized dUTP:dTTP ratio, Uracil-DNA Glycosylase (UDG or UNG), SYBR Green, and Fluorescein Passive Reference Dye in a proprietary reaction buffer. The hot start Taq polymerase has no polymerase activity prior to the initial heat activation step, which allows reaction assembly at room temperature, as well as higher specificity and sensitivity. Since the mix contains dUTP and UDG, carryover contamination prevention can be performed prior to amplification. VeriQuest Fast SYBR Green qPCR Master Mix with Fluorescein offers the same sensitivity and specificity found with our standard mode mixes producing results in a fraction of the time.
VeriQuest Fast SYBR Green qPCR Master Mix with Fluorescein shows exceptional efficiency and high specificity on challenging templates such as high GC- and AT-rich regions. The optimized formulation ensures no sacrifice in quality for increased speed. Performance comparisons highlight the sensitiv-ity and reliable performance of this solution.The sensitivity of the master mix allows for discrimination from a 1.33-fold difference in gene target amount detected. In Fig. 2, a 1.33 to 10-fold dilution series of 10 ng to 1 ng of cDNA reverse-transcribed from HeLa total RNA was amplified with an efficiency of >99%.
VeriQuest Fast SYBR Green qPCR Master Mix with Fluorescein is highly stable and easy to work with. It remains stable at room temperature for 72 hours in
a pre-assembled reaction and can be stored at 4°C for convenient handling. The mix also allows for room temperature reaction set-up. The speed and stability make VeriQuest Fast SYBR Green qPCR Master Mix with Fluorescein ideal for high-through-put handling.
Components:100 reactions 1 ml500 reactions 5 ml1,000 reactions 2 x 5 ml2,500 reactions 5 x 5 ml5,000 reactions 10 x 5 ml
Storage conditionsStore at -20°C for long-term storage. Store at 4-8°C for short-term storage (≤3 months).
PCR Tools | Real-Time qPCR
Fig. 2. High sensitivity and precision in limited target quantification. Amplification plot (left) and standard curve (right) from real-time PCR for a 1.33 to 10-fold dilution series of 10 ng to 1 ng of cDNAs reverse-transcribed from HeLa total RNA.
GAPDH Kras
10 43 2 1 ng RNA
10 4 3 2 1 ng RNA
GAPDH2y = -3.3773x + 22.224
EPCR = 97.7%R² = 0.9955
Krasy = -3.2461x + 30.591
EPCR = 103.3%R² = 0.9993
151719212325272931
-0.2 0 0.2 0.4 0.6 0.8 1 1.2
Ct
log input
GAPDH2 Kras
Fig.1. Linear detection range of VeriQuest Fast SYBR Green qPCR Master Mix. Amplification plot and standard curve from real-time PCR for a dilution series of a synthetic target with starting amounts of 109 copies amplified in four replicate reactions using the ABI 7500 Real-Time PCR System and GAPDH primers in fast mode with 5 minutes activation at 95°C. The amplification process was lin-ear over eight orders of magnitude.
15
20
25
30
35
-4.0 -2.0 0.0 2.0
Ct
log ng input
B2M
ABI Fast SYBR
Bio-Rad iTaq Fast
VeriQuest Fast SYBR
VeriQuest Fast SYBRy = -3.1802 + 23.205EPCR = 106.3%R2 = 0.9999
Bio-Rad iTaq Fast SYBRy = -3.3764 + 22.908EPCR = 97.8%R2 = 0.9991
ABI Fast SYBRy = -3.1984 + 23.755EPCR = 105.4%R2 = 0.9995
Fig. 3. Exceptional results with performance comparison. Standard curves from real-time PCR for a 10-fold dilution series of 10 ng to 1 pg cDNA reverse-transcribed from HeLa total RNA amplified in duplicate reactions with VeriQuest Fast SYBR Green qPCR Master Mix, Bio-Rad iTaq Fast SYBR Green Supermix with ROX™, and ABI Fast SYBR Green Master Mix.
VeriQuest Fast SYBRy = -3.1802 + 23.205EPCR = 106.3%R2 = 0.9999
Bio-Rad iTaq Fast SYBRy = -3.3764 + 22.908EPCR = 97.8%R2 = 0.9991
ABI Fast SYBRy = -3.1984 + 23.755EPCR = 105.4%R2 = 0.9995
Fig. 3. Exceptional results with performance comparison. Standard curves from real-time PCR for a 10-fold dilution series of 10 ng to 1 pg cDNA reverse-transcribed from HeLa total RNA amplified in duplicate reactions with VeriQuest Fast SYBR Green qPCR Master Mix, Bio-Rad iTaq Fast SYBR Green Supermix with ROX™, and ABI Fast SYBR Green Master Mix.
32 888-362-2447 | 216-765-5000 | usb.affymetrix.com
75762 100 reactions 500 reactionsCustoms by request
HotStart-IT SYBR Green qPCR Master Mix uses a novel hot start method developed at Affymetrix called primer sequestration. With this method, an oligo binding protein sequesters primers at lower temperatures making them unavailable for use by Taq DNA Polymerase. Following the initial denatur-ation step, the binding protein is inactivated and the primers are released. HotStart-IT SYBR Green qPCR Master Mix is supplied as a 2X pre-mixed formula-tion containing HotStart-IT Taq DNA Polymerase, MgCl2, Ultrapure nucleotides, and SYBR Green I for use in real-time quantitative PCR reactions (qPCR). Simply add DNA template, primers, and water and the reactions are ready for cycling. Separate tubes of the passive reference dyes ROX (for ABI, Stratagene, and other ROX instruments) and fluorescein (for BioRad and other instruments) are included for added convenience.
The master mix has SYBR Green I Dye which detects any double-stranded DNA that accumulates during the amplification process. For carry-over prevention methods, use HotStart-IT SYBR Green PCR Master Mix with UDG (2X), PN 75760 or VeriQuest SYBR Green qPCR Master Mix (2X), PN 75600.
Two passive reference dyesThe SYBR Green I Dye is compatible with any instrument’s standard SYBR filter set (typically, FAM setting) and the separate ROX and Fluorescein Passive Reference Dyes allow normalization of well-to-well variations that may occur independent of the reactions (e.g., pipetting errors, detection system limitations, etc.).
StableRepeated freeze-thaw cycles have no observed effect on performance.
HotStart-IT SYBR Green qPCR Master Mix (2X):The mix combines HotStart-IT Taq DNA Polymerase, SYBR Green I, MgCl2, and Ultrapure nucleotides in a unique buffer formulation. Magnesium and nucleotide concentrations are 5 mM and 0.4 mM each, respectively.
Components:HotStart-IT SYBR Green qPCR Master Mix (2X): 100 reactions 2 x 1.25 ml 500 reactions 12.5 ml25 mM MgCl2ROX Passive Reference DyeFluorescein Passive Reference Dye
Shipping and storage:Shipped on dry ice. Store at -20°C. Mix well prior to use.
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First-Strand cDNA Synthesis Kit for Real-Time PCR75780 50 reactions
AMV Reverse Transcriptase70041Y 200 units70041Z 1,000 units
M-MLV Reverse Transcriptase78306 25,000 units 100,000 units
Water, Nuclease-Free71786 10 x 1 ml 100 ml 500 ml 1 L 5 L
HotStart-IT SYBR Green qPCR Master Mix (2X)
PCR Tools | Real-Time qPCR
75760 100 reactions 500 reactionsCustoms by request
HotStart-IT SYBR Green qPCR Master Mix with Uracil-DNA Glycosylase (UDG or UNG) uses a novel hot start method developed at Affymetrix called primer sequestration. With this method, a protein binds and sequesters primers at lower temperatures making them unavailable for use by Taq DNA Polymerase. Following the initial denaturation step, the protein is inactivated and the primers are released. HotStart-IT SYBR Green qPCR Master Mix with UDG is supplied as a 2X pre-mixed formulation containing HotStart-IT Taq DNA Polymerase, MgCl2, Ultrapure nucleotides with an optimized dUTP to dTTP ratio, heat-labile UDG, and SYBR Green I for use in real-time quantitative PCR reactions (qPCR). Simply add DNA template, primers, and water and the reactions are ready for cycling. Separate tubes of passive reference dyes ROX (for ABI, Stratagene, and other ROX instruments) and fluorescein (for BioRad and other instruments) are included for added convenience.
Since the mix contains dUTP and UDG, carry-over contamination prevention can be performed. A heat-labile version of UDG that can be irreversibly heat-inactivated is used instead of E. coli UDG, which has been shown to exhibit residual activity following PCR reactions(1). The SYBR Green I Dye detects any double-stranded DNA that accumulates during the amplification process. The hot start feature enhances SYBR-based qPCR reactions by reducing primer-dimer formation which increases specificity and sensitivity. HotStart-IT SYBR Green qPCR Master Mix with UDG has excellent sensitivity as it detects fewer than 10 target copies, performs over a linear dynamic range of 7 to 8 orders of mag-nitude, and is compatible with a variety of real-time PCR instruments.
Multiple platform compatibilityThe SYBR Green I Dye is compatible with any instru-ment’s standard SYBR filter (typically, FAM setting) and the separate ROX and Fluorescein Passive Reference Dyes allow normalization of well-to-well variations that may occur independent of the reactions (e.g., pipetting errors, detection system limitations, etc.).
StableRepeated freeze-thaw cycles have no observed effect on performance.
HotStart-IT SYBR Green qPCR Master Mix with UDG Formulation (2X):The mix combines HotStart-IT Taq DNA Polymerase, heat-labile UDG, SYBR Green I, MgCl2, and Ultrapure nucleotides with an optimized dUTP to dTTP ratio in a unique buffer formulation. Magne-sium and nucleotide concentrations are 5 mM and 0.4 mM each, respectively.
Components:HotStart-IT SYBR Green qPCR Master Mix with
UDG (2X) 100 reactions 2 x 1.25 ml 500 reactions 12.5 ml25 mM MgCl2ROX Passive Reference DyeFluorescein Passive Reference Dye
Shipping and storage:Shipped on dry ice. Store at -20°C. Mix well prior to use.
References:1. Thornton, C. G., Hartley, J. L., and Rashtchian, A.
(1992) BioTechniques 13, 180-184.2. Longo, M. C., Berninger, M. S., and Hartley, J. L.
(1990) Gene 93, 125-128.
HotStart-IT SYBR Green qPCR Master Mix with UDG (2X)
33For bulk or alternate pack sizes, email us at [email protected].
PCR Tools | Real-Time qPCR
75650 (50 µl reaction volume) 40 reactions (1 ml) 200 reactions (5 ml) 400 reactions (2 x 5 ml) 1,000 reactions (5 x 5 ml) 2,000 reactions (10 x 5 ml)
VeriQuest Probe qPCR Master Mix is supplied as a 2X pre-mixed formulation containing both ROX and UDG in an optimized buffer for quality results in real-time quantitative PCR assays. The proprietary reaction buffer with optimum MgCl2 concentration is specifically designed for robust probe hybridiza-tion and efficient cleavage of TaqMan probes. Simply add DNA template, primers, probe(s), and water and the reactions are ready for cycling. The master mix contains a chemically-modified Taq DNA Polymerase, MgCl2, ultrapure nucleotides with an optimized dUTP:dTTP ratio, Uracil-DNA Glycosylase (UDG or UNG), and ROX Passive Reference Dye in a proprietary reaction buffer.
Features:�� One-tube master mix with ROX Reference Dye included�� Similar protocols, same probes, and same primers for standard mode amplification�� Consistency over a broad dynamic range: 9 orders of magnitude linear detection range with a correlation coefficient = 0.999�� Exceptional performance with challenging GC-rich regions �� Detect 2 copies of target from genomic DNA and differentiate less than a two-fold difference.�� Contains dUTP and Uracil-DNA Glycosylase (UDG or UNG) for optional carryover contamination prevention.�� Hot start PCR for room temperature reaction setup�� Stable at room temperature for 72 hours in a preassembled reaction�� Validated for use with a variety of 5’ reporter fluorophores and 3’ quenchers, as well as TaqMan MGB containing probes
Reproducible, consistent results while main-taining precision and efficiencyThe convenience of a master mix formulation with-out sacrificing quality. VeriQuest Probe qPCR Master Mix provides consistency over a broad dynamic template concentration range allowing 9 orders of magnitude linear detection (Fig. 1 and 2). High preci-sion in target quantification and discrimination of a 1.33 to 10-fold dilution series with VeriQuest Probe qPCR Master Mix ensure accurate results (Fig. 3).
Enhanced amplification sensitivity and accuracy of low copy number detectionReliable detection from as little as 2 copies of target from genomic DNA and differentiation from less than a two-fold difference (Fig. 3).
Exceptional performance with challenging templatesEven in high GC and AT rich regions VeriQuest Probe qPCR Master Mix offers exceptional performance and high specificity so you have the confidence to verify your gene expression results (Fig. 4).
Ready-to-use, one-tube replacementThe one-tube master mix contains all neces-sary components; chemically-modified Taq DNA Polymerase, MgCl2, ultrapure nucleotides with an optimized dUTP:dTTP ratio, Uracil-DNA Glycosylase (UDG or UNG), and ROX as the pas-sive reference dye, all premixed in a proprietary reaction buffer. A drop-in replacement for use on any leading PCR platform that utilizes ROX.
Passive Reference Dye and UDG includedROX Passive Reference Dye is an inert dye, whose fluorescence does not change during the reaction. This signal normalization is necessary to correct for well-to-well differences that may occur due to artifacts such as pipetting errors or sample evapora-tion. Uracil-DNA Glycosylase (UDG or UNG) and dUTP offer an option for carry-over contamination prevention from previous PCR amplifications. All are at optimized levels so there is no need to adjust concentrations.
Highly stable and easy to work withVeriQuest Probe qPCR Master Mix is stable at room temperature for 72 hours in a pre-assembled reaction and can be stored at 4°C for convenient handling and room temperature reaction set up; no lost time waiting for your master mix to thaw (Figs. 5 and 6). Testing of 10 freeze-thaw cycles showed no loss in master mix performance. Ideal for high-throughput handling.
Consistent performance of single-plex and duplex detectionSingleplex (S) and duplex detection (D) of target genes can be performed using VeriQuest Probe qPCR Master Mix, making this ideal for gene expression verification.
Components:40 reactions: 1 ml200 reactions: 5 ml400 reactions: 2 x 5 ml1,000 reactions: 5 x 5 ml2,000 reactions: 10 x 5 mlPCR Qualified Water
References:1. VanGuilder, H. D., Vrana, K. E., and Freeman, W. M.
(2008) BioTechniques 44 (5), 619-626.2. Longo, M. C., Berninger, M. S., and Hartley, J. L.
(1990) Gene 93, 125-128.3. Tewhey, R. et al. (2009) Nature Biotechnology 27,
1025-1031.
VeriQuest Probe qPCR Master Mix (2X)
Fig. 1. Linear detection range of VeriQuest Probe qPCR Master Mix. Real-time amplification plot from a 10-fold dilution series of a GAPDH synthetic target with starting amounts of 1010 copies amplified in four replicate reactions using the ABI 7500 PCR System and GAPDH primer-probe set (FAM-BHQ®-1).
Fig. 2. Inset for linear detection range. Amplification plot from a 10-fold dilution series.
Fig. 3. High precision in target quantification with VeriQuest Probe qPCR Master Mix. Amplification plot from real-time PCR for a 1.33 to 10-fold dilution series of 10 ng to 1 ng of cDNAs reverse-transcribed from HeLa total RNA.
(Continued on next page)
34 888-362-2447 | 216-765-5000 | usb.affymetrix.com
VeriQuest Probe qPCR Master Mix (2X) continued...
Fig. 4. Amplification of GC-rich target with VeriQuest Probe qPCR Master Mix. qPCR standard curves for a 10-fold dilution series of 10 ng to 10 pg human male genomic DNA amplified in four replicate reactions with VeriQuest Probe qPCR Master Mix, ABI TaqMan® Gene Expression Master Mix and ABI TaqMan Universal Master Mix II using the ABI 7500 PCR System for detection of a 71.2% GC-target amplicon (PROC, NT_022135.16)(3).
Fig. 6. Inset defining Ct levels of 1 ng and 10 pg of cDNAs reverse transcribed from HeLa total RNA. GAPDH was detect-ed from preassembled PCR reactions incubated at room temperature for 48 and 72 hours.
Fig. 5. VeriQuest Probe qPCR Master Mix stability for reli-able performance of high-throughput handling. Pre-assembled PCR reaction stability after 48 and 72 hour incubations at room temperature from quantification of GAPDH cDNAs.
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VeriQuest Probe qPCR Master Mix, No Reference Dye (2X)75660 40 reactions 200 reactions 400 reactions 1,000 reactions 2,000 reactions
VeriQuest Fast Probe qPCR Master Mix (2X)75680 100 reactions 500 reactions 1,000 reactions 2,500 reactions 5,000 reactions
VeriQuest Fast Probe qPCR Master Mix, No Reference Dye (2X)75685 100 reactions 500 reactions 1,000 reactions 2,500 reactions 5,000 reactions
First-Strand cDNA Synthesis Kit for Real-Time PCR75780 50 reactions
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PCR Tools | Real-Time qPCR
35For bulk or alternate pack sizes, email us at [email protected].
75660 (50 µl reaction volume) 40 reactions (1 ml) 200 reactions (5 ml) 400 reactions (2 x 5 ml) 1,000 reactions (5 x 5 ml) 2,000 reactions (10 x 5 ml)
VeriQuest Probe qPCR Master Mix, No Reference Dye is supplied as a 2X pre-mixed formulation con-taining UDG in an optimized buffer for quality results in real-time quantitative PCR assays. The proprietary reaction buffer with optimum MgCl2 concentration is specifically designed for robust probe hybridization and efficient cleavage of TaqMan probes. Simply add DNA template, primers, probe(s), and water, and the reactions are ready for cycling.
Features:�� Enhanced amplification sensitivity and accuracy of low copy number detection�� Reproducible, consistent results while maintaining precision and efficiency�� Exceptional performance with challenging templates�� Ready-to-use, one-tube replacement�� UDG included�� Highly stable and easy to work with�� Ample detection of up to 4 different target genes
Enhanced sensitivity on amplification for low copy number detectionReliable detection from as little as two copies of target from genomic DNA and differentiation from less than a two-fold difference.
Reproducible, consistent results while main-taining precision and efficiencyThe convenience of a master mix formulation with-out sacrificing quality. VeriQuest Probe qPCR Master Mix, No Reference Dye provides consistency over a broad dynamic template concentration range, al-lowing 9 orders of magnitude linear detection. High precision in target quantification and discrimination of a 1.33 to 10-fold dilution series with VeriQuest Probe qPCR Master Mix, No Reference Dye ensures accurate results.
Exceptional performance with challenging templatesEven in high GC and AT rich regions, VeriQuest Probe qPCR Master Mix, No Reference Dye offers excep-tional performance and high specificity so you have the confidence to verify your gene expression results.
Ready-to-use, one-tube replacementThe one-tube master mix contains all necessary com-ponents: chemically-modified Taq DNA Polymerase, MgCl2, ultrapure nucleotides with an optimized dUTP:dTTP ratio, and Uracil-DNA Glycosylase (UDG or UNG) in a proprietary reaction buffer. Simply add your DNA template, probe, primers, and water, and you can begin your qPCR reaction. For use in your new or existing protocols, on any leading PCR plat-form that does not require a passive reference dye, providing easy transition from other master mixes. Examples of PCR platforms include: Eppendorf Mastercycler®, Helixis Pixo™, Qiagen Rotor-Gene™ Q, Roche LightCycler® 480/1536, TaKaRa TP-800, Bio-Rad CFX96/CFX384, Bio-Rad Opticon 2™, Bio-Rad Chromo 4™, Cepheid Smart Cycler®, and Corbett Rotor-Gene™.
UDG included for carry-over contamination preventionUracil-DNA Glycosylase (UDG or UNG) and dUTP offer an option for carry-over contamination preven-tion from previous PCR amplifications. Both are at optimized levels so there is no need to adjust concentrations.
Highly stable and easy to work withVeriQuest Probe qPCR Master Mix, No Reference Dye is stable at room temperature for 72 hours in a preassembled reaction. This mix can be stored at 4°C for convenient handling and room temperature reaction set up, with no time lost waiting for your master mix to thaw. Testing of 10 freeze thaw cycles showed no loss in master mix performance, making it ideal for high-throughput handling.
Ample detection of up to 2 different target genesSingle-plex (S) and duplex detection (D) of target genes is possible with VeriQuest Probe qPCR Master Mix, No Reference Dye for gene expression verification.
Components:40 reactions: 1 ml200 reactions: 5 ml400 reactions: 2 x 5 ml1,000 reactions: 5 x 5 ml2,000 reactions: 10 x 5 mlPCR Qualified Water
Storage:Shipped on dry ice. Store at -20°C for long-term storage. Store at 4-8°C for short-term storage (≤3 months).
VeriQuest Probe qPCR Master Mix, No Reference Dye (2X)
Real-time amplification plot and standard curve from a 10-fold dilution series of a synthetic target with starting amounts of 1010 copies ampli-fied in four replicate reactions using the Bio-Rad MyiQ Real-Time PCR System and GAPDH primer-probe set (Fam-BHQ®-1).
Linear detection range of VeriQuest Probe qPCR Master Mix, No Reference Dye
PCR Tools | Real-Time qPCR
36 888-362-2447 | 216-765-5000 | usb.affymetrix.com
VeriQuest Fast Probe qPCR Master Mix (2X)
75680 (20 µl reaction volume) 100 reactions (1 ml) 500 reactions (5 ml) 1,000 reactions (2 x 5 ml) 2,500 reactions (5 x 5 ml) 5,000 reactions (10 x 5 ml)
Features:�� Uses fast mode thermal cycling conditions for results in one-third of the time when compared to standard run protocols.�� One-tube master mix with ROX Reference Dye included – just add template, primers and water �� Exceptional performance with challenging GC-rich regions�� Contains dUTP and Uracil-DNA Glycosylase (UDG or UNG) for carryover contamination prevention.�� Hot start PCR for room temperature reaction setup�� Sensitivity and precision with limited targets
VeriQuest Fast Probe qPCR Master Mix (2X) is a ready-to-use master mix optimized for TaqMan® probe detection on all instruments using fast mode cycling protocols that utilize ROX passive reference dye. The 2X ready-to-use master mix contains hot start VeriQuest Taq DNA Polymerase, MgCl2, ultrapure nucleotides with an optimized dUTP:dTTP ratio, Uracil-DNA Glycosylase (UDG or UNG), and ROX Passive Reference Dye in a proprietary reaction buffer. The hot start Taq polymerase has no polymerase activity prior to the initial heat activation step, which allows reaction assembly at room temperature as well as higher specificity and sensitivity. Since the mix contains dUTP and UDG, carryover contamination prevention can be performed prior to amplification. VeriQuest Fast Probe qPCR Master Mix offers the
same sensitivity and specificity found with our stan-dard mode mixes with results in a fraction of the time.
VeriQuest Fast Probe qPCR Master Mix shows excep-tional efficiency and high specificity on challenging templates such as high GC- and AT-rich regions. The optimized formulation ensures no sacrifice in quality for increased speed. Performance comparisons highlight the sensitivity and reliable performance of this platform-independent solution.The sensitivity of the master mix allows for discrimination from a 1.33-fold difference in gene target amount detected. In Fig. 1, a 1.33 to 10-fold dilution series of 10 ng to 1 ng of cDNA reverse-transcribed from HeLa total RNA was amplified with an efficiency of >98.8%.
VeriQuest Fast Probe qPCR Master Mix is highly stable and easy to work with. It remains stable at
room temperature for 72 hours in a pre-assembled reaction and can be stored at 4°C for convenient handling. The mix also allows for room tempera-ture reaction set-up. The speed and stability make VeriQuest Fast Probe qPCR Master Mix ideal for high-throughput handling.
Components:100 reactions 1 ml500 reactions 5 ml1,000 reactions 2 x 5 ml2,500 reactions 5 x 5 ml5,000 reactions 10 x 5 ml
Storage conditionsStore at -20°C for long-term storage. Store at 4-8°C for short-term storage (≤3 months).
Linear detection range of VeriQuest Fast Probe qPCR Master Mix. Amplification plot and standard curve from real-time PCR for a dilution series of a synthetic target with starting amounts of 1010 cop-ies amplified in four replicate reactions using the ABI 7500 Real-Time PCR System and GAPDH primers in fast mode with 5 minutes activa-tion at 95°C. The amplification process was linear over eight orders of magnitude.
PCR Tools | Real-Time qPCR
Fig. 2. High sensitivity and precision in limited target quantification. Amplification plot (bottom) and standard curve (top) from real-time PCR for a 1.33 to 10-fold dilution series of 10 ng to 1 ng of cDNA reverse-transcribed from HeLa total RNA.
Cyclophilin
10 4 3 2 1 ng RNA
Kras
10 4 3 2 1 ng RNA
Cyclophiliny = -3.3841x + 24.142
EPCR = 97.5%R² = 0.9977
Krasy = -3.3502x + 31.829
EPCR = 98.8%R² = 0.9973
20
22
24
26
28
30
32
34
-0.2 0 0.2 0.4 0.6 0.8 1 1.2
Ct
log input
Cyclophilin Kras
Fig. 3. VeriQuest Fast Probe qPCR Master Mix stability. GAPDH was detected from pre-assembled PCR reactions incubated at room temperature for 72 hours. Results were compared to the freshly prepared mix (0 hours).
14
19
24
29
34
39
-3.0 -2.0 -1.0 0.0 1.0 2.0 3.0
Ct
0 hours 72 hours
0 hours y = -3.4119x + 24.224
EPCR = 96.7%R² = 0.9995
72 hours y = -3.3581x + 24.134
EPCR= 98.5%R² = 0.9996
37For bulk or alternate pack sizes, email us at [email protected].
75685 (20 µl reaction volume) 100 reactions (1 ml) 500 reactions (5 ml) 1,000 reactions (2 x 5 ml) 2,500 reactions (5 x 5 ml) 5,000 reactions (10 x 5 ml)
Features:�� Uses fast mode thermal cycling conditions for results in one-third of the time when compared to standard run protocols.�� One-tube master mix – just add template, primers, and water �� Exceptional performance with challenging GC-rich regions�� Contains dUTP and Uracil-DNA Glycosylase (UDG or UNG) for carryover contamination prevention.�� Hot start PCR for room temperature reaction setup�� Sensitivity and precision with limited targets
VeriQuest Fast Probe qPCR Master Mix, No Reference Dye (2X) is a ready-to-use master mix optimized for TaqMan® probe detection on all instruments using fast mode cycling protocols that do not require use of a passive reference dye. The 2X ready-to-use master mix contains hot start VeriQuest Taq DNA Polymerase, MgCl2, ultrapure nucleotides with an optimized dUTP:dTTP ratio, and Uracil-DNA Glycosylase (UDG or UNG) in a proprietary reaction buffer. The hot start Taq polymerase has no polymerase activity prior to the initial heat activation step, which allows reaction assembly at room tem-perature as well as higher specificity and sensitivity. Since the mix contains dUTP and UDG, carryover contamination prevention can be performed prior to amplification. VeriQuest Fast Probe qPCR Master
Mix, No Reference Dye offers the same sensitivity and specificity found with our standard mode mixes with results in a fraction of the time.
VeriQuest Fast Probe qPCR Master Mix, No Refer-ence Dye shows exceptional efficiency and high specificity on challenging templates such as high GC- and AT-rich regions The optimized formulation ensures no sacrifice in quality for increased speed. Performance comparisons highlight the sensitivity and reliable performance of this platform-indepen-dent solution.The sensitivity of the master mix allows for discrimination from a 1.33-fold difference in gene target amount detected. In Fig. 1, a 1.33 to 10-fold dilution series of 10 ng to 1 ng of cDNA reverse-transcribed from HeLa total RNA was ampli-fied with an efficiency of >98.8%. VeriQuest Fast Probe qPCR Master Mix, No Reference Dye is highly
stable and easy to work with. It remains stable at room temperature for 72 hours in a pre-assembled reaction and can be stored at 4°C for convenient handling. The mix also allows for room temperature reaction set-up. The speed and stability make Veri-Quest Fast Probe qPCR Master Mix, No Reference Dye ideal for high-throughput handling.
Components:100 reactions 1 ml500 reactions 5 ml1,000 reactions 2 x 5 ml2,500 reactions 5 x 5 ml5,000 reactions 10 x 5 ml
Storage conditionsStore at -20°C for long-term storage. Store at 4-8°C for short-term storage (≤3 months).
VeriQuest Fast Probe qPCR Master Mix, No Reference Dye (2X)
PCR Tools | Real-Time qPCR
Linear detection range of VeriQuest Fast Probe qPCR Master Mix. Amplification plot and standard curve from real-time PCR for a dilution series of a synthetic target with starting amounts of 1010 cop-ies amplified in four replicate reactions using the ABI 7500 Real-Time PCR System and GAPDH primers in fast mode with 5 minutes activa-tion at 95°C. The amplification process was linear over eight orders of magnitude.
Fig. 2. High sensitivity and precision in limited target quantification. Amplification plot (bottom) and standard curve (top) from real-time PCR for a 1.33 to 10-fold dilution series of 10 ng to 1 ng of cDNA reverse-transcribed from HeLa total RNA.
Cyclophilin
10 4 3 2 1 ng RNA
Kras
10 4 3 2 1 ng RNA
Cyclophiliny = -3.3841x + 24.142
EPCR = 97.5%R² = 0.9977
Krasy = -3.3502x + 31.829
EPCR = 98.8%R² = 0.9973
20
22
24
26
28
30
32
34
-0.2 0 0.2 0.4 0.6 0.8 1 1.2
Ct
log input
Cyclophilin Kras
Fig. 3. VeriQuest Fast Probe qPCR Master Mix stability. GAPDH was detected from pre-assembled PCR reactions incubated at room temperature for 72 hours. Results were compared to the freshly prepared mix (0 hours).
14
19
24
29
34
39
-3.0 -2.0 -1.0 0.0 1.0 2.0 3.0
Ct
0 hours 72 hours
0 hours y = -3.4119x + 24.224
EPCR = 96.7%R² = 0.9995
72 hours y = -3.3581x + 24.134
EPCR= 98.5%R² = 0.9996
38 888-362-2447 | 216-765-5000 | usb.affymetrix.com
75766 100 reactions 500 reactionsCustoms by request
HotStart-IT Probe qPCR Master Mix uses a novel hot start method developed at Affymetrix called primer sequestration. With this method, a protein binds and sequesters primers at lower temperatures mak-ing them unavailable for use by Taq DNA Polym-erase. Following the initial denaturation step, the protein is inactivated and the primers are released. HotStart-IT Probe qPCR Master Mix is supplied as a 2X pre-mixed formulation containing HotStart-IT Taq DNA Polymerase, MgCl2, and Ultrapure nucleo-tides for use in real-time quantitative PCR reactions (qPCR) with fluorescent probes. Simply add DNA template, primers, probe(s), and water and the reac-tions are ready for cycling. A separate tube of ROX passive reference dye (for ABI, Stratagene, and other instruments) is included for added convenience.
The master mix is formulated for use with fluo-rescent probes such as TaqMan Probes, Molecular Beacons, and others(1-2). Since fluorescent probes are designed to hybridize to the target of interest, detection specificity is greatly increased relative to non-specific dsDNA binding dyes such as SYBR Green I. The Taq DNA Polymerase used in the master mix has the 5'→3' exonuclease activity necessary for efficient removal of the 5'-fluorophore from the 3'-quencher in TaqMan probes. HotStart-IT Probe qPCR Master Mix has excellent sensitivity as it detects fewer than 10 target copies, performs over a broad, linear dynamic range of 7 to 8 orders of magnitude, and is compatible with a variety of real-time PCR instruments. The mix does not have dUTP in place of dTTP and is incompatible with carry-over contamination prevention methods using Uracil-DNA Glycosylase (UDG or UNG). For carry-over prevention methods, use HotStart-IT Probe qPCR Master Mix with UDG (2X), PN 75764 or VeriQuest Probe qPCR Master Mix (2X), PN 75650.
StableRepeated freeze-thaw cycles have no observed effect on performance.
HotStart-IT Probe qPCR Master Mix (2X):The mix combines HotStart-IT Taq DNA Polymerase (with 5'→3' exonuclease activity), MgCl2, and Ultrapure nucleotides in a unique buffer formulation. Magnesium and nucleotide concentrations are 6 mM and 0.4 mM each, respectively.
Components:HotStart-IT Probe qPCR Master Mix (2X) 100 reactions 2 x 1.25 ml 500 reactions 12.5 ml25 mM MgCl2ROX Passive Reference Dye
Shipping and storage:Shipped on dry ice. Store at -20°C. Mix well prior to use.
References:1. Livak, K. J., Flood, S. J., Marmaro, J., Giusti, W., and
Deetz, K. (1995) PCR Methods Appl. 4, 357-362.2. Tyagi, S., and Kramer, F. R. (1996) Nat. Biotechnol.
14, 303-308.
HotStart-IT Probe qPCR Master Mix (2X)
75764 100 reactions 500 reactionsCustoms by request
HotStart-IT Probe qPCR Master Mix with Uracil-DNA Glycosylase (UDG or UNG) uses a novel hot start method developed at Affymetrix called primer sequestration. With this method, a protein binds and sequesters primers at lower temperatures making them unavailable for use by Taq DNA Polymerase. Following the initial denaturation step, the protein is inactivated and the primers are released. HotStart-IT Probe qPCR Master Mix with UDG is supplied as a 2X pre-mixed formulation containing HotStart-IT Taq DNA Polymerase, MgCl2, Ultrapure nucleotides with an optimized dUTP to dTTP ratio, and heat-labile UDG for use in real-time quantitative PCR reactions (qPCR) with fluorescent probes. Simply add DNA template, primers, probe(s), and water and the reac-tions are ready for cycling. A separate tube of ROX passive reference dye (for ABI, Stratagene, and other instruments) is included for added convenience.
Since the mix contains dUTP and UDG, carry-over contamination prevention can be performed. A heat-labile version of UDG that can be irreversibly heat-inactivated is used instead of E. coli UDG, which has been shown to exhibit residual activity
following PCR reactions(1). The mix is formulated for use with fluorescent probes such as TaqMan Probes, Molecular Beacons, and others(3-4). Since fluorescent probes are designed to hybridize to the target of interest, detection specificity is greatly increased relative to non-specific dsDNA binding dyes such as SYBR Green I. The Taq DNA Polymerase used in the master mix has the 5'→3' exonuclease activity necessary for efficient removal of the 5'-fluorophore from the 3'-quencher in TaqMan probes. HotStart-IT Probe qPCR Master Mix with UDG has excellent sensitivity as it detects fewer than 10 target copies, performs over a broad, linear dynamic range of 7 to 8 orders of magnitude, and is compatible with a vari-ety of real-time PCR instruments.
Multiple platform compatibilitySpecialized buffer with optimal MgCl2 concentration and dUTP to dTTP ratio performs well on a variety of platforms. Also, the separate tube of ROX Passive Reference Dye allows normalization of well-to-well variations that may occur independent of the reactions (e.g., pipetting errors, detection system limitations, etc.).
StableRepeated freeze-thaw cycles have no observed effect on performance.
HotStart-IT Probe qPCR Master Mix with UDG (2X):The mix combines HotStart-IT Taq DNA Polymerase (with 5'→3' exonuclease activity), heat-labile UDG, MgCl2, and Ultrapure nucleotides with an optimized dUTP to dTTP ratio in a unique buffer formulation. Magnesium and nucleotide concentrations are 6 mM and 0.4 mM each, respectively.
Components:HotStart-IT Probe qPCR Master Mix with UDG (2X) 100 reactions 2 x 1.25 ml 500 reactions 12.5 ml25 mM MgCl2ROX Passive Reference Dye
Shipping and storage:Shipped on dry ice. Store at -20°C. Mix well prior to use.
References:1. Thornton, C. G., Hartley, J. L., and Rashtchian, A.
(1992) BioTechniques 13, 180-184.2. Longo, M. C., Berninger, M. S., and Hartley, J. L.
(1990) Gene 93, 125-128.3. Livak, K. J., Flood, S. J., Marmaro, J., Giusti, W., and
Deetz, K. (1995) PCR Methods Appl. 4, 357-362.4. Tyagi, S., and Kramer, F. R. (1996) Nat. Biotechnol.
14, 303-308.
HotStart-IT Probe qPCR Master Mix with UDG (2X)
PCR Tools | Real-Time qPCR
39For bulk or alternate pack sizes, email us at [email protected].
75780 50 reactions (20 µl)
The First-Strand cDNA Synthesis Kit for Real-Time PCR is optimized for reverse transcription of RNA and produces first-strand cDNA template suitable for real-time PCR.
�� Robust performance: Optimized to reliably generate cDNA from full-length transcripts longer than 12 kb.�� Sensitive: Sensitive and reliable real-time RT-PCR analysis of multiple user-defined targets from as little as 10 pg of starting total RNA.�� Consistent performance: Optimized for generating first-strand cDNA to be used in real-time PCR. HeLa total RNA and primers for qPCR are included as a positive control.�� Versatile: Three different priming strategies are provided to meet experimental needs. Mixed priming strategy overcomes 5' and/or 3' end bias associated with typical oligo(dT)n or random hexamer priming strategies.
Applications:Conversion of RNA into single-stranded cDNA involves a complex, enzyme-catalyzed reaction known as reverse transcription (RT). RT followed by real-time or quantitative PCR (real-time RT-PCR or qPCR) amplification is the most sensitive technique for mRNA detection and quantification currently available(1).
This kit is specifically designed to convert RNA into first-strand cDNA that is suitable for real-time PCR applications.
First-strand cDNA generated by this kit is also suitable for end-point PCR. RT-PCR can be used in a variety of applications, such as qualitative and quantitative analyses of cellular RNAs, characteriza-tion of RNA splice variants, and the generation and cloning of cDNAs.
Mixed priming strategy eliminates end-biasThis kit includes an optimized Primer Mix which results in the generation of first-strand cDNAs from an entire transcript without the end-bias observed with typical oligo(dT)n or random hexamer primers (Fig. 1). This mixed primer strategy overcomes variability in real-time PCR gene expression analysis that can result from using different individual primers(2). For convenience, anchored oligo(dT)23VN and random hexamer primers are also included.
Robust and sensitive performanceThis kit has been designed to routinely generate cDNAs from transcripts longer than 12 kb (Fig. 2). Consistent results are obtained using any of the three priming strategies provided with this kit. Reverse transcription can be performed with as little as 10 pg of total RNA, allowing sensitive down-stream analysis (Fig. 3).
Kit components:10X RT Buffer10 mM dNTPsRNase Inhibitor (10 units/µl)RNase-Free, DEPC-Treated WaterM-MLV RTAnchored Oligo(dT)23VN (50 µM)Random Hexamers (75 µM )10X Primer MixHeLa Total RNA (100 ng/µl)Control Primer Mix for qPCR
References:1. Bustin, S., Benes, V., Nolan, T., and Pfaffl, M.
(2005) J. Mol. Endocrinology 34, 597-601.2. Ståhlberg, A., Håkansson, J., Xian, X., Semb, H.,
and Kubista, M. (2004) Clinical Chem. 50(3), 509-515.
First-Strand cDNA Synthesis Kit for Real-Time PCR
Fig. 1. The Primer Mix solution greatly reduces bias for sequences near the 5' and/or 3' ends of cDNAs produced. The β-glucuronidase (GUSB) mRNA was reverse transcribed from 100 ng of HeLa cell total RNA using (A) Random Hexamers, (B) Anchored Oligo(dT)23VN, or (C) the Primer Mix. Amplicons located near the 5' end (red) or 3' end (green) of the GUSB transcript were amplified by real-time PCR (on an ABI 7500 Fast instrument) using 1 µl of each reverse transcription reaction in 20 µl real-time PCR reactions (in duplicate) and HotStart-IT SYBR Green qPCR Master Mix (PN 75762).
Fig. 2. Assay flexibility and consistent priming strategies. The First-Strand cDNA Synthesis Kit for Real-Time PCR has been opti-mized for assay flexibility in carrying out reverse transcription using various priming strategies and diverse RNA templates. (A) Clathrin (NM_004859) and (B) Utrophin (NM_007124) amplicons were amplified using gene specific primers designed for real-time PCR (brown arrows). Reverse transcription was performed using HeLa cell total RNA (100 ng) and three different priming strategies: Anchored Oligo dT23VN primer (red), Random Hexamers (blue), and Primer Mix (green). 1 µl of each reverse transcription reaction was then used in 20 µl real-time PCR reactions on an ABI 7500 Fast instrument using HotStart-IT SYBR Green qPCR Master Mix (PN 75762).
Fig. 3. Assay sensitivity and dynamic range. Various amounts of HeLa cell total RNA, ranging from 850 ng down to 10 pg, were reverse transcribed using anchored oligo(dT)23VN as the primer. 1 µl of each reverse transcription reaction was then used in 20 µl real-time PCR reactions (in duplicate) to amplify a 122 bp GAPDH amplicon (on an ABI 7500 Fast instrument), using HotStart-IT SYBR Green qPCR Master Mix (PN 75762). GAPDH amplification and linear correlation curve above show the wide dynamic range and sensitivity of the First-Strand cDNA Synthesis Kit for Real-Time PCR.
PCR Tools | Real-Time Reverse Transcription PCR (qRT-PCR)
40 888-362-2447 | 216-765-5000 | usb.affymetrix.com
VeriQuest SYBR Green One-Step qRT-PCR Master Mix (2X)
75705 (50 µl reaction volume) 40 reactions (1 ml) 200 reactions (5 ml)
Features:�� Reproducible, consistent results while maintaining precision and efficiency�� Multiple platform compatibility with all real-time master mixes�� One-step, sequential reaction format for minimum hands on time�� Pre-mixed format with VeriQuest qRT-PCR mixes reduces potential contamination�� Exceptional stability even after 10 freeze-thaw cycles
VeriQuest SYBR Green One-Step qRT-PCR Master Mix is a one-step, ready-to-use master mix for RNA quantification with SYBR Green detection. The optimized buffer formulation offers specificity, ro-bustness, and reliability. The RT-PCR process converts and amplifies single-stranded RNA template yielding double-stranded DNA product. One-Step RT-PCR uses gene specific primers, designed to match RNA/cDNA targets, in a single-tube/plate, one-step reac-tion. This approach offers tremendous convenience when applied to analysis of single targets from mul-tiple RNA samples. Also, it minimizes the possibility of introducing contaminants into reactions between the RT and PCR steps, since both steps are carried out sequentially without opening the reaction tubes/plates between the steps(1-5).
Reproducible, consistent results while main-taining precision and efficiencyThis master mix provides the convenience of a master mix formulation without sacrificing quality. VeriQuest SYBR Green One-Step qRT-PCR Master Mix exhibits excellent sensitivity, as it can detect fewer than 10 target copies and performs over a broad, linear dynamic range of 8 orders of magni-tude (Figs. 1 and 2).
Multiple platform compatibility with all real-time master mixesThis master mix contains ROX passive reference dye in an optimized concentration for use on all instru-ments that require ROX dye for signal normalization. ROX Passive Reference Dye is an inert dye, whose fluorescence does not change during the reaction. This signal normalization is necessary to correct for well-to-well differences that may occur due to arti-facts such as pipetting errors or sample evaporation. VeriQuest SYBR Green OneStep qRT-PCR Master Mix is also available for instruments that require fluorescein. To determine which master mix is appro-priate for your thermal cycler, refer to our instrument compatibility guide.
Pre-mixed format with VeriQuest qRT-PCR mixes reduces potential contaminationThe 2X master mix contains 100X RT Enzyme Mix and 2X qRT-PCR Mix for RNA quantification with SYBR Green. The 100X RT Enzyme Mix is a blend of reverse transcriptase and RNase Inhibitor. The 2X qRT-PCR Mix contains hot start VeriQuest Taq DNA Polymerase, ultrapure nucleotides, SYBR Green I and ROX Passive Reference Dye in an optimized buffer formulation for quantitative, real-time PCR detection with SYBR Green.
Exceptional stability even after 10 freeze-thaw cyclesVeriQuest SYBR Green One-Step qRT-PCR Master Mix is stable and easy to work with. Performance testing of 10 freeze thaw cycles showed no loss in master mix performance, making this ideal for high-throughput handling.
Components:100X RT Enzyme Mix for SYBR Green Assay2X qRT-PCR Mix with SYBR Green 40 reactions 1 ml200 reactions 5 ml
Storage conditionsStore at -20°C for long-term storage. Store at 4-8°C for short-term storage (≤3 months).
References:1. Goblet, C., Prost, E., and Whalen, R. G. (1989)
Nucleic Acids Res. 17, 2144.2. Sambrook, J. and Russell, D. W. (2001) “Molecular
Cloning: A Laboratory Manual,” Cold Spring Harbor Laboratory Press, 8.46-8.53.
3. Sellner, L. N., Coelen, R. J., and MacKenzie, J. S. (1992) Nucleic Acids Res. 20, 1487-1490.
4. Roth, M. J., Tanese, N., and Goff, S. P. (1985) J. Biol. Chem. 260, 9326-9335.
5. Saiki, R. K., Gelfand, D. H., Stoffel, S., Scharf, S. J., Higuchi, R., Horn, G. T., Mullis, K. B., and Erlich, H. A. (1988) Science 239, 487-491.
Fig. 2. Melt Curve analysis for VeriQuest SYBR Green One-Step qRT-PCR Master Mix.
Fig. 1. Consistent results over an extremely broad dynamic range. Linear detection range of VeriQuest SYBR Green One-Step qRT-PCR Master Mix. RNA from human muscle cells was reverse tran-scribed and amplified using 18S primers. Triplicate reactions were run on an ABI 7500 Real-Time PCR System
PCR Tools | Real-Time Reverse Transcription PCR (qRT-PCR)
41For bulk or alternate pack sizes, email us at [email protected].
VeriQuest SYBR Green One-Step qRT-PCR Master Mix with Fluorescein (2X)
75715 (50 µl reaction volume) 40 reactions (1 ml) 200 reactions (5 ml)
Features:�� Reproducible, consistent results while maintaining precision and efficiency�� Multiple platform compatibility with all real-time master mixes�� One-step, sequential reaction format for minimum hands on time�� Pre-mixed format with VeriQuest qRT-PCR mixes reduces potential contamination�� Exceptional stability even after 10 freeze-thaw cycles
VeriQuest SYBR Green One-Step qRT-PCR Master Mix with Fluorescein is a one-step, ready-to-use master mix for RNA quantification with SYBR Green detection. The optimized buffer formulation offers specificity, robustness, and reliability. The RT-PCR process converts and amplifies single-stranded RNA template yielding double-stranded DNA product. One-Step RT-PCR uses gene specific primers, de-signed to match RNA/cDNA targets, in a single-tube/plate, one-step reaction.
This approach offers tremendous convenience when applied to analysis of single targets from multiple RNA samples. Also, it minimizes the possibility of introducing contaminants into reactions between the RT and PCR steps, since both steps are carried out sequentially without opening the reaction tubes/plates between the steps(1-5).
Reproducible, consistent results while main-taining precision and efficiencyThis master mix provides the convenience of a master mix formulation without sacrificing quality. VeriQuest SYBR Green One-Step qRT-PCR Master Mix with Fluorescein exhibits excellent sensitivity, as it can detect fewer than 10 target copies and performs over a broad, linear dynamic range of 8 orders of magnitude (Figs. 1 and 2).
Multiple platform compatibility with all real-time master mixesThis master mix contains fluorescein passive refer-ence dye in an optimized concentration for use on instruments that require fluorescein dye for signal normalization (i.e., BioRad iCycler, MyiQ, iQ5). Fluo-rescein Passive Reference Dye is an inert dye, whose fluorescence does not change during the reaction. This signal normalization is necessary to correct for well-to-well differences that may occur due to arti-facts such as pipetting errors or sample evaporation. VeriQuest SYBR Green One-Step qRT-PCR Master Mix is also available for instruments that require ROX reference dye. To determine which master mix is appropriate for your thermal cycler, refer to our instrument compatibility guide.
Pre-mixed format with VeriQuest qRT-PCR mixes reduces potential contaminationThe 2X master mix contains 100X RT Enzyme Mix and 2X qRT-PCR Mix for RNA quantification with SYBR Green. The 100X RT Enzyme Mix is a blend of reverse transcriptase and RNase Inhibitor. The 2X qRT-PCR Mix contains chemically-modified VeriQuest Taq DNA Polymerase, ultrapure nucleotides, SYBR Green I, and Fluorescein Passive Reference Dye in optimized buffer formulation for quantitative, real-time PCR detection with SYBR Green.
Exceptional stability even after 10 freeze-thaw cyclesVeriQuest SYBR Green One-Step qRT-PCR Master Mix with Fluorescein is stable and easy to work with. Performance testing of 10 freeze thaw cycles showed no loss in master mix performance, making this ideal for high-throughput handling.
Components:100X RT Enzyme Mix for SYBR Green Assay2X qRT-PCR Mix with SYBR Green 40 reactions 1 ml200 reactions 5 ml
Storage conditionsStore at -20°C for long-term storage. Store at 4-8°C for short-term storage (≤3 months).
References:1. Goblet, C., Prost, E., and Whalen, R. G. (1989)
Nucleic Acids Res. 17, 2144.2. Sambrook, J. and Russell, D. W. (2001) “Molecular
Cloning: A Laboratory Manual,” Cold Spring Harbor Laboratory Press, 8.46-8.53.
3. Sellner, L. N., Coelen, R. J., and MacKenzie, J. S. (1992) Nucleic Acids Res. 20, 1487-1490.
4. Roth, M. J., Tanese, N., and Goff, S. P. (1985) J. Biol. Chem. 260, 9326-9335.
5. Saiki, R. K., Gelfand, D. H., Stoffel, S., Scharf, S. J., Higuchi, R., Horn, G. T., Mullis, K. B., and Erlich, H. A. (1988) Science 239, 487-491.
PCR Tools | Real-Time Reverse Transcription PCR (qRT-PCR)
Fig. 2. Melt Curve analysis for VeriQuest SYBR Green One-Step qRT-PCR Master Mix.
Fig. 1. Consistent results over an extremely broad dynamic range. Linear detection range of VeriQuest SYBR Green One-Step qRT-PCR Master Mix. RNA from human muscle cells was reverse tran-scribed and amplified using 18S primers. Triplicate reactions were run on an ABI 7500 Real-Time PCR System
42 888-362-2447 | 216-765-5000 | usb.affymetrix.com
VeriQuest Probe One-Step qRT-PCR Master Mix (2X)
75700 (50 µl reaction volume) 40 reactions (1 ml) 200 reactions (5 ml)
Features:�� Reproducible, consistent results while maintaining precision and efficiency�� Multiple platform compatibility with all real-time master mixes�� One-step, sequential reaction format for minimum hands on time�� Easily perform analysis of single or duplex targets from multiple RNA samples�� Pre-mixed format with VeriQuest qRT-PCR mixes reduces potential contamination�� Exceptional stability even after 10 freeze-thaw cycles
VeriQuest Probe One-Step qRT-PCR Master Mix is a one-tube, one-step, ready-to-use master mix for RNA quantification with TaqMan® probe detection. The optimized buffer formulation offers specificity, ro-bustness, and reliability. The RT-PCR process converts and amplifies single-stranded RNA template yielding double-stranded DNA product. One-Step RT-PCR uses gene specific primers, designed to match RNA/cDNA targets, in a single-tube/plate, one-step reac-tion. This approach offers tremendous convenience when applied to analysis of single targets from mul-tiple RNA samples. Also, it minimizes the possibility of introducing contaminants into reactions between the RT and PCR steps, since both steps are carried out sequentially without opening the reaction tubes/plates between the steps(1-5).
Reproducible, consistent results while main-taining precision and efficiencyThis master mix provides the convenience of a master mix formulation without sacrificing quality. VeriQuest Probe One-Step qRT-PCR Master Mix ex-hibits excellent sensitivity, as it can detect fewer than 10 target copies and performs over a broad, linear dynamic range of 8 orders of magnitude (Fig. 1).
Multiple platform compatibility with all real-time master mixesThis master mix contains ROX passive reference dye in an optimized concentration for use on all instru-ments that require ROX dye for signal normalization. ROX Passive Reference Dye is an inert dye, whose fluorescence does not change during the reaction. This signal normalization is necessary to correct for well-to-well differences that may occur due to arti-facts such as pipetting errors or sample evaporation.
Pre-mixed format with VeriQuest qRT-PCR mixes reduces potential contaminationThe 2X master mix contains reverse transcriptase, RNase inhibitor, our chemically modified VeriQuest Taq DNA polymerase, ROX passive reference dye, and ultrapure nucleotides all in an optimized buffer composition. Simply add your RNA template, probe, primers, and water, and you can begin your qRT-PCR reaction. For use in your new or existing protocols, on any leading real-time qPCR platform that utilizes ROX.
Exceptional stability even after 10 freeze-thaw cyclesVeriQuest Probe One-Step qRT-PCR Master Mix is stable and easy to work with. Performance testing of 10 freeze thaw cycles showed no loss in master mix performance, making this ideal for high-throughput handling.
Components:40 reactions 1 ml200 reactions 5 ml
Storage conditionsStore at -20°C for long-term storage. Store at 4-8°C for short-term storage (≤3 months).
References:1. Goblet, C., Prost, E., and Whalen, R. G. (1989)
Nucleic Acids Res. 17, 2144. 2. Sambrook, J. and Russell, D. W. (2001) “Molecular
Cloning: A Laboratory Manual,” Cold Spring Harbor Laboratory Press, 8.46-8.53.
3. Sellner, L. N., Coelen, R. J., and MacKenzie, J. S. (1992) Nucleic Acids Res. 20, 1487-1490.
4. Roth, M. J., Tanese, N., and Goff, S. P. (1985) J. Biol. Chem. 260, 9326-9335.
5. Saiki, R. K., Gelfand, D. H., Stoffel, S., Scharf, S. J., Higuchi, R., Horn, G. T., Mullis, K. B., and Erlich, H. A. (1988) Science 239, 487-491.
PCR Tools | Real-Time Reverse Transcription PCR (qRT-PCR)
Fig. 1. Consistent results over an extremely broad dynamic range. Linear detection range of VeriQuest Probe One-Step qRT-PCR Master Mix. RNA from human muscle cells was reverse transcribed and amplified using 18S primers. Triplicate reactions were run on an ABI 7500 Real-Time PCR System
43For bulk or alternate pack sizes, email us at [email protected].
VeriQuest Probe One-Step qRT-PCR Master Mix, No Reference Dye (2X)
75710 (50 µl reaction volume) 40 reactions (1 ml) 200 reactions (5 ml)
Features:�� Reproducible, consistent results while maintaining precision and efficiency�� Multiple platform compatibility with all real-time master mixes�� One-step, sequential reaction format for minimum hands on time�� Easily perform analysis of single or duplex targets from multiple RNA samples�� Pre-mixed format with VeriQuest qRT-PCR mixes reduces potential contamination�� Exceptional stability even after 10 freeze-thaw cycles
VeriQuest Probe One-Step qRT-PCR Master Mix, No Reference Dye is a one-tube, one-step, ready-to-use master mix for RNA quantification with TaqMan® probe detection. The optimized buffer formulation of-fers specificity, robustness, and reliability. The RT-PCR process converts and amplifies single-stranded RNA template yielding double-stranded DNA product.
One-Step RT-PCR uses gene specific primers, designed to match RNA/cDNA targets, in a single-tube/plate, one-step reaction. This approach offers tremendous convenience when applied to analysis of single targets from multiple RNA samples. Also, it minimizes the possibility of introducing contami-nants into reactions between the RT and PCR steps, since both steps are carried out sequentially without opening the reaction tubes/plates between the steps(1-5).
Reproducible, consistent results while main-taining precision and efficiencyThis master mix provides the convenience of a master mix formulation without sacrificing quality. VeriQuest Probe One-Step qRT-PCR Master Mix, No Reference Dye exhibits excellent sensitivity, as it can detect fewer than 10 target copies and performs over a broad, linear dynamic range of 8 orders of magnitude (Fig. 1). Product comparisons show opti-mal results in head-to-head evaluations (Fig. 2).
Multiple platform compatibility with all real-time master mixesThis master mix does not contain a passive reference dye, making it useful for thermo cyclers that do not
require the use of a dye for signal normalization (i.e., LightCycler®, RotorGene, Opticon®). To determine which master mix is appropriate for your thermal cycler, refer to our instrument compatibility guide.
Easily perform analysis of single, duplex, or triplex targets from multiple RNA samplesDuplex detection of target RNA using VeriQuest Probe One-Step qRT-PCR Master Mix, No Reference Dye can be performed with the same sensitivity and specificity as single-plex reactions.
Pre-mixed format with VeriQuest qRT-PCR mixes reduces potential contaminationThe 2X master mix contains reverse transcriptase, RNase inhibitor, our chemically modified VeriQuest Taq DNA Polymerase, and ultrapure nucleotides all in an optimized buffer composition. Simply add your RNA template, probe, primers, and water, and you can begin your qRT-PCR reaction. For use in your new or existing protocols, or on any leading real-time qPCR platform that does not require the use of a reference dye.
Exceptional stability even after 10 freeze-thaw cyclesVeriQuest Probe One-Step qRT-PCR Master Mix, No Reference Dye is stable and easy to work with. Per-formance testing of 10 freeze thaw cycles showed no loss in master mix performance making this ideal for high-throughput handling.
Components:40 reactions 1 ml200 reactions 5 ml
Storage conditionsStore at -20°C for long-term storage. Store at 4-8°C for short-term storage (≤3 months).
References:1. Goblet, C., Prost, E., and Whalen, R. G. (1989)
Nucleic Acids Res. 17, 2144.2. Sambrook, J. and Russell, D. W. (2001) “Molecular
Cloning: A Laboratory Manual,” Cold Spring Harbor Laboratory Press, 8.46-8.53.
3. Sellner, L. N., Coelen, R. J., and MacKenzie, J. S. (1992) Nucleic Acids Res. 20, 1487-1490.
4. Roth, M. J., Tanese, N., and Goff, S. P. (1985) J. Biol. Chem. 260, 9326-9335.
5. Saiki, R. K., Gelfand, D. H., Stoffel, S., Scharf, S. J., Higuchi, R., Horn, G. T., Mullis, K. B., and Erlich, H. A. (1988) Science 239, 487-491.
PCR Tools | Real-Time Reverse Transcription PCR (qRT-PCR)
Fig. 1. Consistent results over an extremely broad dynamic range. Linear detection range of VeriQuest Probe One-Step qRT-PCR Master Mix. RNA from human muscle cells was reverse transcribed and amplified using 18S primers. Triplicate reactions were run on an ABI 7500 Real-Time PCR System
13.0
19.8 23.4
26.8
30.4 33.2
14.0
20.9
24.4 27.8
30.7 32.9
13.3
20.0
23.6 26.9
30.5 33.5
0
5
10
15
20
25
30
35
40
100 ng 10 ng 1 ng 100 pg 10 pg 1 pg 100 fg
Ct
HeLa Total RNA
ABI TaqMan Fast Virus 1-Step
ABI TaqMan RNA-to-CT™ 1-Step
VeriQuest One-Step Probe
16.5 17.5 16.6
slope EPCR R2
ABI TaqMan Fast Virus 1-Step -3.4087 96.50% 0.9994
ABI TaqMan RNA-to-CT 1-Step -3.2142 104.70% 0.9953
VeriQuest One-Step Probe -3.4057 96.62% 0.9998
Fig. 2. Exceptional results with performance comparisons. Product comparison—VeriQuest Probe One-Step qRT-PCR Master Mix. Ct values, R2, and PCR efficiency from real-time PCR for a 10-fold dilution series of 100 ng to 100 fg HeLa total RNA amplified in duplicate reac-tions with VeriQuest Probe One-Step qRT-PCR Master Mix, ABI TaqMan Fast Virus 1-Step Master Mix, and ABI TaqMan RNA-to-CT 1-Step Kit using the ABI 7500 Fast Real-Time PCR System and GAPDH primers and probe (Fam-BHQ) in standard mode with 15 minute RT reaction at 50°C.
44 888-362-2447 | 216-765-5000 | usb.affymetrix.com
75770 100 reactions 500 reactionsCustoms by request
Description:HotStart-IT SYBR Green One-Step qRT-PCR Master Mix Kit provides optimal performance and maximum convenience for real-time, quantitative analysis of RNA templates in a single reaction format. The RT-PCR process converts and amplifies single-stranded RNA template yielding double-stranded DNA product. One-step RT-PCR uses gene specific primers, designed to match RNA/cDNA targets, in a single-tube/plate, one-step reaction. This approach offers tremendous convenience when applied to analysis of single targets from multiple RNA samples. Also, it minimizes the possibility of introducing con-taminants into reactions between the RT and PCR steps, since both steps are carried out sequentially without opening the reaction tubes/plates between the steps(1-5).
HotStart-IT SYBR Green One-Step qRT-PCR Master Mix Kit includes M-MLV RT, RNase Inhibitor, and a 2X Master Mix containing HotStart-IT Taq DNA Polymerase, MgCl2, Ultrapure nucleotides, and SYBR Green I in an optimized reaction buffer. HotStart-IT SYBR Green qPCR Master Mix uses a novel hot
start method developed at Affymetrix called primer sequestration. This novel hot start feature increases the specificity and sensitivity of SYBR-based qRT-PCR reactions by substantially reducing primer-dimer formation. With this method, the HotStart-IT protein binds and sequesters primers at lower tempera-tures making them unavailable for use by Taq DNA Polymerase. Following reverse transcription and the subsequent heat denaturation step, the primer binding protein is inactivated and the primers are released.
This kit exhibits excellent sensitivity as it can detect fewer than 10 target copies, performs over a broad, linear dynamic range of 6 to 7 orders of magnitude, and is compatible with most real-time PCR instru-ments.
Based on Affymetrix primer sequestration technology�� Increases specificity and prevents primer-dimer formation�� No DNA template damage – no extensive heating step needed to denature hotstart component
Shipping and storage:Shipped on dry ice. Store at -20°C. Mix all compo-nents well prior to use. Light sensitive components should be protected from excessive light exposure.
Components: 100 reaction kit 500 reaction kit Rctn. Vol. = 50 µl Rctn. Vol. = 50 µlHotStart-IT SYBR Green
qPCR Master Mix (2X) 2 x 1.25 ml 1 x 12.5 ml25 mM MgCl2 1 x 1 ml 5 x 1 mlROX Passive Reference Dye 1 x 100 μl 1 x 500 μlFluorescein Passive
Reference Dye 1 x 100 μl 1 x 500 μlM-MLV RT 1 x 40 μl 1 x 200 μlRNase Inhibitor (10 units/μl) 1 x 40 μl 1 x 200 μlRNase-Free Water,
DEPC-Treated 3 x 1 ml 1 x 15 ml
References:1. Goblet, C., Prost, E., and Whalen, R. G. (1989)
Nucleic Acids Res. 17, 2144.2. Sambrook, J. and Russell, D. W. (2001) “Molecular
Cloning: A Laboratory Manual,” Cold Spring Harbor Laboratory Press, 8.46-8.53.
3. Sellner, L. N., Coelen, R. J., and MacKenzie, J. S. (1992) Nucleic Acids Res. 20, 1487-1490.
4. Roth, M. J., Tanese, N., and Goff, S. P. (1985) J. Biol. Chem. 260, 9326-9335.
5. Saiki, R. K., Gelfand, D. H., Stoffel, S., Scharf, S. J., Higuchi, R., Horn, G. T., Mullis, K. B., and Erlich, H. A. (1988) Science 239, 487-491.
HotStart-IT SYBR Green One-Step qRT-PCR Master Mix Kit
75772 100 reactions 500 reactionsCustoms by request
Description:HotStart-IT Probe One-Step qRT-PCR Master Mix Kit provides optimal performance and maximum convenience for real-time, quantitative analysis of RNA templates in a single reaction format. The RT-PCR process converts and amplifies single-stranded RNA template yielding double-stranded DNA product. One-Step RT-PCR uses gene specific primers, designed to match RNA/cDNA targets, in a single-tube/plate, one-step reaction. This approach offers tremendous convenience when applied to analysis of single targets from multiple RNA samples. This minimizes the possibility of introducing con-taminants into reactions between the RT and PCR steps, since both steps are carried out sequentially without opening the reaction tubes/plates between the steps(1-5).
HotStart-IT Probe One-Step qRT-PCR Master Mix Kit includes M-MLV RT, RNase Inhibitor, and a 2X Mas-ter Mix containing HotStart-IT Taq DNA Polymerase, MgCl2, and Ultrapure nucleotides in an optimized reaction buffer for use with fluorescent probes. HotStart-IT Probe qPCR Master Mix uses a novel hot start method developed at Affymetrix called primer
sequestration (see PN 71195). With this method, the HotStart-IT protein binds and sequesters primers at lower temperatures making them unavailable for use by Taq DNA Polymerase. Following reverse transcription and the subsequent heat denaturation step, the primer binding protein is inactivated and the primers are released.
This kit is formulated for use with fluorescent probes such as TaqMan Probes, Molecular Beacons, and others(6-7). Since fluorescent probes are designed to hybridize to the target of interest, detection specificity is greatly increased relative to non-specific dsDNA binding dyes such as SYBR Green I. The Taq DNA Polymerase used in this master mix has the 5' to 3' exonuclease activity necessary for efficient removal of the 5'-fluorophore from the 3'-quencher in TaqMan probes.
Based on Affymetrix primer sequestration technology�� Increases specificity and prevents primer-dimer formation�� No DNA template damage – no extensive heating step needed to denature hotstart component
Shipping and storage:Shipped on dry ice. Store at -20°C. Mix all compo-nents well prior to use. Light sensitive components should be protected from excessive light exposure.
Components: 100 reaction kit 500 reaction kit Rctn. Vol. = 50 µl Rctn. Vol. = 50 µl
HotStart-IT Probe qPCR Master Mix (2X) 2 x 1.25 ml 1 x 12.5 ml
25 mM MgCl2 1 x 1 ml 5 x 1 mlROX Passive Reference Dye 1 x 100 μl 1 x 500 μlM-MLV RT 1 x 40 μl 1 x 200 μlRNase Inhibitor (10 units/μl) 1 x 40 μl 1 x 200 μlRNase-Free Water,
DEPC-Treated 3 x 1 ml 1 x 15 ml
References:1. Goblet, C., Prost, E., and Whalen, R. G. (1989)
Nucleic Acids Res. 17, 2144.2. Sambrook, J. and Russell, D. W. (2001) “Molecular
Cloning: A Laboratory Manual,” Cold Spring Harbor Laboratory Press, 8.46-8.53.
3. Sellner, L. N., Coelen, R. J., and MacKenzie, J. S. (1992) Nucleic Acids Res. 20, 1487-1490.
4. Roth, M. J., Tanese, N., and Goff, S. P. (1985) J. Biol. Chem. 260, 9326-9335.
5. Saiki, R. K., Gelfand, D. H., Stoffel, S., Scharf, S. J., Higuchi, R., Horn, G. T., Mullis, K. B., and Erlich, H. A. (1988) Science 239, 487-491.
6. Livak, K. J., Flood, S. J., Marmaro, J., Giusti, W., and Deetz, K. (1995) PCR Methods Appl. 4, 357-362.
7. Tyagi, S., and Kramer, F. R. (1996) Nat. Biotechnol. 14, 303-308.
HotStart-IT Probe One-Step qRT-PCR Master Mix Kit
PCR Tools | Real-Time Reverse Transcription PCR (qRT-PCR)
45For bulk or alternate pack sizes, email us at [email protected].
78370 100 reactions
RT-PCR Master Mix provides maximum con-venience and optimal performance for highly sensitive and specific one-step RT-PCR reactions. This unique formulation combines all the reagents necessary for successful RT-PCR. Simply add RT-PCR Master Mix to RNA template, primers, and RNase-free water, and the reactions are ready to begin.
During reverse transcription-polymerase chain reac-tion (RT-PCR), reverse transcriptase converts RNA template to cDNA which is subsequently amplified by a thermostable DNA polymerase(1,2). One-step RT-PCR is a variation of RT-PCR in which the com-ponents necessary for both the RT and PCR steps are combined in a single-tube and the reactions are performed sequentially(2). This single-step, closed-tube approach simplifies the expression analyses of one or a few genes from multiple RNA samples and reduces the risk of contaminating samples. RT-PCR Master Mix further simplifies this technique by providing one-step RT-PCR in a pre-mixed format.
ConvenientRT-PCR Master Mix saves time and reduces potential contamination errors by eliminating several pipetting steps. Since the mix is pre-formulated and thoroughly QC tested, experimental variability is significantly reduced. This translates into greater reproducibility in demanding, high-throughput experiments. For a 50 µl reaction, simply add 25 µl of RT-PCR Master Mix to primers, RNA template, and RNase-free H2O.
Improve specificity and sensitivityAmplify RT-PCR products from less template, with lower background, and with little or no optimization. Targets may be routinely detected from just 100 fg of total RNA. For example, human β-actin is detected from 100 femtograms of total RNA, which represents about 1/100th of the total RNA of a single human cell (Fig. 1).
Optimal formulationAn enhanced buffer allows for RT reaction tempera-tures up to 50°C, which can improve detection of more difficult targets. This is because higher RT tem-peratures reduce non-specific priming and facilitate melting of RNA secondary structure(3). In addition, the mix has the wild-type, RNase H-plus form of M-MLV Reverse Transcriptase which has been shown to improve the sensitivity for certain targets(4). The RNase H activity degrades the RNA part of the cDNA/RNA hybrid following reverse transcription which may prevent its inhibition during subsequent PCR steps. RT-PCR Master Mix has been specifically designed to detect targets whose sizes are generally less than 1.0 kb. For targets greater than 1.0 kb and for high fidelity, use FideliTaq RT-PCR Master Mix (PN 71185).
StableRT-PCR Master Mix withstands repeated freeze-thaw cycles with no observed decrease in performance (Fig. 2).
Functional test:Tested by amplifying a 459 bp β-actin target from 10 pg of human placental total RNA.
RT-PCR Master Mix Formulation (2X):Includes M-MLV Reverse Transcriptase, Taq DNA Polymerase, recombinant RNase Inhibitor, nucleo-tides, and magnesium in a novel RT-PCR buffer. Magnesium concentration is 3 mM in the 2X RT-PCR Master Mix.
Components:RT-PCR Master Mix is supplied in kit form with the following components sufficient for 100 reactions in a 50 µl reaction volume:
4 x 625 µl RT-PCR Master Mix (2X)3 x 1 ml RNase-free Water1 x 1 ml 25 mM MgCl2 Solution
Shipping and storage:Shipped on dry ice. Store at -20°C. Mix well prior to use.
References:1. Sambrook, J. and Russell, D. W. (2001) “Molecular
Cloning: A Laboratory Manual,” Cold Spring Harbor Laboratory Press, 8.46-8.53.
2. Sellner, L. N., Coelen, R. J., and Mackenzie, J. S. (1992) Nucl. Acids Res. 20, 1487-1490.
3. Malboeuf, C. M., Isaacs, S. J., Tran, N. H., and Kim, B. (2001) BioTechniques 30, 1074-1084.
4. Polumuri, S. K., Ruknudin, A., and Schulze, D. H. (2002) BioTechniques 32, 1224-1225.
RT-PCR Master Mix (2X)
-RNA
1 kb –
500 bp –
200 bp –
M -RT 10 pg1 pg 100 fg
– β-actin (459 bp)
Total RNA
M -RNA -RT pre post pre post pre post pre post
10 ng 1 ng 100 pg 10 pg Total RNA
1 kb –500 bp –
200 bp –
– numb (455 bp)
M -RNA -RT pre post pre post pre post pre post
1 ng 100 pg 10 pg 1 pg Total RNA
1 kb –500 bp –
200 bp –– β-actin (459 bp)
Fig. 1. Sensitivity of the RT-PCR Master Mix. A 459 bp frag-ment of the human β-actin gene was RT-PCR amplified from the indicated amounts of human placental total RNA. RT temperature was 50°C and primers were at 0.8 µM. The -RNA control included no RNA in the sample and the -RT control included 100 ng of total RNA but used the Taq PCR Master Mix (PN 71162, without reverse transcriptase) to test for any contaminating genomic DNA in the RNA sample. “M” is the DNA marker lane. Since the typical human cell contains about 10 picograms of total RNA, this target’s detection at 100 femtograms represents about 1/100th of the total RNA of a typical human cell.
Fig. 2. Stability of the RT-PCR Master Mix. Targets were RT-PCR amplified from the indicated amounts of human placental total RNA. RT temperature was 50°C and primers were at 0.8 µM. The -RNA control included no RNA in the sample and the -RT control includ-ed 100 ng of total RNA but used the Taq PCR Master Mix (PN 71162, without reverse tran-scriptase) to test for any contaminating genomic DNA in the RNA sample. “Pre” indicates lanes in which the Master Mix was not freeze-thawed and “Post” indicates lanes in which the Master Mix was freeze-thawed 15 times, alternating between dry ice and a room tem-perature water bath. “M” is the DNA marker lane.
PCR Tools | Reverse Transcription RT-PCR—One-Step Master Mix
46 888-362-2447 | 216-765-5000 | usb.affymetrix.com
71185 100 reactions
FideliTaq RT-PCR Master Mix provides maximum convenience and optimal performance for highly sensitive, specific, and accurate one-step RT-PCR reactions. This unique formulation combines all the reagents necessary for successful, high-fidelity RT-PCR. Simply add FideliTaq RT-PCR Master Mix to RNA template, primers, and RNase-free water, and the reactions are ready to begin.
During reverse transcription-polymerase chain reaction (RT-PCR), reverse transcriptase converts RNA template to cDNA which is subsequently amplified by a thermostable DNA polymerase(1,2). One-step RT-PCR is a variation of RT-PCR in which the components necessary for both the RT and PCR steps are combined in a single tube and the reactions are performed sequentially(2). This single-step, closed-tube approach simplifies the expression analyses of one or a few genes from multiple RNA samples and reduces the risk of contaminating samples. FideliTaq RT-PCR Master Mix further simplifies this technique by providing high-fidelity, one-step RT-PCR in a pre-mixed format.
ConvenientFideliTaq RT-PCR Master Mix saves time and reduces potential contamination errors by eliminating several pipetting steps. Since the mix is pre-formulated and thoroughly QC tested, experimental variability is significantly reduced. This translates into greater reproducibility in demanding, high-throughput experiments. For a 50 µl reaction, simply add 25 µl of FideliTaq RT-PCR Master Mix to primers, RNA template, and RNase-free H2O.
High fidelity and longer RT-PCR productsBy incorporating FideliTaq Polymerase into the mix, amplification fidelity is increased up to 6 times over Taq Polymerase alone, which is ideal for cloning applications(3-6). FideliTaq RT-PCR Master Mix gener-ates products with both blunt-ends and those with non-template added adenine on the 3' end. The ratio of blunt-ends to non-template, adenylated-ends varies from primer to primer and base identity at the 3' end(7). Thus, RT-PCR products may be cloned into both blunt-end and TA vectors, although TA vectors may yield more transformants. In addition, longer product sizes can be generated up to about 6 kb (Fig. 1).
Improve specificity and sensitivityAmplify RT-PCR products from less template, with lower background, and with little or no optimization. Targets may be routinely detected from just 100 fg of total RNA. For example, human β-actin is detected from 100 femtograms of total RNA, which represents about 1/100th of the total RNA of a single human cell.
Optimal formulationAn enhanced buffer allows for RT reaction tem-peratures up to 50°C, which can improve detection of more difficult targets. This is because higher RT temperatures reduce non-specific priming and facilitate melting of RNA secondary structure(8). In addition, the mix has the wild-type, RNase H-plus form of M-MLV Reverse Transcriptase which has been shown to improve the sensitivity for certain targets(9). The RNase H activity degrades the RNA part of the cDNA/RNA hybrid following reverse transcription which may prevent its inhibition during subsequent PCR steps.
Stable performanceThe FideliTaq RT-PCR Master Mix withstands repeat-ed freeze-thaw cycles with no observed decrease in performance (Fig. 1).
Functional test:Tested by amplifying a 459 bp β-actin target from 10 pg and a 1.5 kb β-actin target from 100 pg of human placental total RNA, plus a 5.6 kb Clathrin target from 10 ng of pooled human total RNA.
FideliTaq RT-PCR Master Mix Formulation (2X):Includes M-MLV Reverse Transcriptase, FideliTaq DNA Polymerase, recombinant RNase Inhibitor, nucleotides, and magnesium in a novel RT-PCR buffer. Magnesium concentration is 3 mM in the 2X FideliTaq RT-PCR Master Mix.
Components:FideliTaq RT-PCR Master Mix is supplied in kit form with the following components sufficient for 100 reactions in a 50 µl reaction volume:
4 x 625 µl FideliTaq RT-PCR Master Mix (2X)3 x 1 ml RNase-free Water1 x 1 ml 25 mM MgCl2 Solution
Shipping and storage:Shipped on dry ice. Store at -20°C. Mix well prior to use.
References:1. Sambrook, J. and Russell, D. W. (2001) “Molecular
Cloning: A Laboratory Manual,” Cold Spring Harbor Laboratory Press, 8.46-8.53.
2. Sellner, L. N., Coelen, R. J., and Mackenzie, J. S. (1992) Nucl. Acids Res. 20, 1487-1490.
3. Barnes, W. M. (1994) Proc. Natl. Acad. Sci. USA 91, 2216-2220.
4. Cheng, S., Fockler, C., Barnes, W. M., and Higuchi, R. (1994) Proc. Natl. Acad. Sci. USA 91, 5695-5699.
5. Barnes, W. M. (1992) Gene 112, 29-35.6. Cline, J., Braman, J. C., and Hogrefe, H. H. (1996)
Nucl. Acids Res. 24, 3546-3551.7. Magnuson, V. L., Ally, D. S., Nylund, S. J.,
Karanjawala, Z. E., Rayman, J. B., Knapp, J. I., Lowe, A. L., Ghosh, S., and Collins, F. S. (1996) BioTechniques 4, 700-709.
8. Malboeuf, C. M., Isaacs, S. J., Tran, N. H., and Kim, B. (2001) BioTechniques 30, 1074-1084.
9. Polumuri, S. K., Ruknudin, A., and Schulze, D. H. (2002) BioTechniques 32, 1224-1225.
FideliTaq RT-PCR Master Mix (2X)
M -RNA -RT pre post pre post pre post pre post
10 ng 1 ng 100 pg 10 pg Total RNA
3 kb –
1.65 kb –1 kb –
– β-actin (1.5 kb)
M -RNA -RT pre post pre post pre post
100 ng 10 ng 1 ng Total RNA
12 kb –
3 kb –1.65 kb –
– Clathrin (5.6 kb)
Fig. 1. Stability and product length associated with the FideliTaq RT-PCR Master Mix. Targets were RT-PCR amplified from the indicated amounts of human total RNA. RT temperature was 50°C and primers were at 0.8 µM. The -RNA control included no RNA in the sample and the -RT control included 100 ng of total RNA but used the Taq PCR Master Mix (PN 71162, without reverse transcriptase) to test for any contaminating genomic DNA in the RNA sample. “Pre” indicates lanes in which the Master Mix was not freeze-thawed and “Post” indicates lanes in which the Master Mix was freeze-thawed 15 times alternating between dry ice and a room temperature water bath. “M” is the DNA marker lane. Human placental RNA was used for actin, and pooled RNA from several human cell-lines was used for Clathrin. The mix withstands repeated freeze-thaw cycles (up to 15) with no effect on product yield and is able to generate targets at least up to 5.6 kb.
PCR Tools | Reverse Transcription RT-PCR—One-Step Master Mix
47For bulk or alternate pack sizes, email us at [email protected].
78350 50 reactions
The One-Step RT-PCR Kit is designed for simple RT-PCR in a one-tube format.�� Streamlines and optimizes the RT and PCR steps�� Provides a starting point for analysis of new RNA targets
Reverse transcription-polymerase chain reaction, or RT-PCR, is a method for converting and amplifying a single-stranded RNA template to yield abundant double-stranded complementary DNA (cDNA) product(1,2). In the RT step, the RT enzyme reverse transcribes an RNA template, yielding single-stranded cDNA. In the PCR step, a thermostable DNA polymerase amplifies the single-stranded cDNA to yield double-stranded cDNA product. One step RT-PCR is a variation on RT-PCR in which all reaction components are mixed in one tube prior to starting the reactions(2). This approach offers simplicity and convenience and minimizes the possibility for contamination.
Complete One-Step RT-PCR KitThe One-Step RT-PCR Kit comes complete and ready-to-use for RT-PCR. The kit can be used with diverse RNA samples and custom primers. The kit uses M-MLV Reverse Transcriptase(3) and Taq DNA Polymerase(4), premixed together at concentrations optimized to balance sensitivity and specificity. An optimized reaction buffer, RNase Inhibitor, Ultrapure dNTPs, supplemental magnesium chloride, and RNase-free water are also included.
Highly sensitive, highly specificThe kit can be used for detection of diverse RNA targets (Fig. 1). Moderately or weakly expressed targets can be detected in 1 ng to 1 µg total RNA or 100 pg to 100 ng polyA RNA. Highly expressed tar-gets can be detected in even lower amounts of RNA (Fig. 2). Standard reaction conditions are sufficient for amplification of most targets. A few simple proto-col adjustments, such as optimization of the amount of primers or addition of supplements for amplifying G+C rich targets, enable specific amplification of other targets(5) (Fig. 3).
Convenient, one-tube formatSetting up reactions is quick and simple, given that the RT and PCR steps are set up simultaneously and then carried out sequentially in one tube. This format eliminates the need to set up RT and PCR indepen-dently, saving time and eliminating a potential point of contamination.
Rapid, reliable resultsThe One-Step RT-PCR Kit is ideal for qualitative analysis of expression of one or a few genes in multiple RNA samples, analysis of specific RNA splice variants, and streamlining the optimization of RT and PCR steps simultaneously. The kit is also particularly good as a starting point for analysis of new RNA targets, given that many targets can be amplified successfully without need for optimization.
Kit components:RT-PCR Enzyme Mix5X RT-PCR Reaction Buffer (includes MgCl2)Ultrapure PCR Nucleotide Mix (10 mM each dATP,
dCTP, dGTP, dTTP)RNase Inhibitor, Recombinant (4 units/µl)25 mM Magnesium ChlorideRNase-Free Water
Shipping and storage:Shipped on dry ice. Store at -20°C.
References:1. Sambrook, J. and Russel, D. W. (2001) “Molecular
Cloning: A Laboratory Manual,” Cold Spring Harbor Laboratory Press, 8.46-8.53.
2. Sellner, L. N., Coelen, R. J., and Mackenzie, J. S. (1992) Nucl. Acids Res. 20, 1487-1490.
3. Roth, M. J., Tanese, N., and Goff, S. P. (1985) J. Biol. Chem. 260, 9326-9335.
4. Saiki, R. K., Gelfand, D. H., Stoffel, S., Scharf, S. J., Higuchi, R., Horn, G. T., Mullis, K. B., and Erlich, H. A. (1988) Science 239, 487-491.
5. Tech Tip 206, Simple Approaches for Optimization of RT-PCR, USB Corporation, Cleveland, Ohio.
One-Step RT-PCR Kit
75780 50 reactions (20 µl)See page 39
First-Strand cDNA Synthesis Kit for Real-Time PCR
Fig. 1. Amplification of diverse RNA targets by one-step RT-PCR. Target (source): β-actin (100 ng total RNA, human liver), Numb (100 ng total RNA, human liver), Ubiquitin (1 µg total RNA, Arabidopsis leaf), and Terminal Deoxynucleotidyl Transferase (TdT) (100 ng polyA RNA, calf thymus). Gene specific primers were designed to generate products of particular sizes. M: Marker; Lanes 1-3: Actin, 0.5 kb, 1.0 kb, 1.5 kb; Lanes 4-5: Numb, 0.5 kb, 1.0 kb; Lane 6: Ubiquitin, 1.5 kb; Lane 7: TdT, 1.5 kb.
0.5 kb –
1 kb –
2 kb –
Fig. 2. Highly sensitive detection of β-actin target from human liver total RNA and polyA RNA, by one-step RT-PCR. Primers were used at 0.8 µM, a rela-tively high concentration, in order to achieve high sensitivity. Total RNA: 1 µg to 1 pg, 10-fold dilutions shown in Lanes 1-7. Poly A RNA: 100 ng to 100 fg, 10-fold dilutions shown in Lanes 1-7.
0.2 kb –
0.5 kb –
1.0 kb –
TotalRNA
PolyARNA
Fig. 3. Flexibility for optimization. For targets with high G+C contents, such as 0.34 kb Notch3 (G+C: 77%), adding supplements and/or increasing the temperature of the reverse transcription step, may improve specificity. Compare results for standard reac-tion conditions versus reaction with betaine supple-ment and elevated temperature.
0.2 kb –
0.5 kb –
1.5 M betaine:Temp (°C):
PCR Tools | Reverse Transcription RT-PCR—Kits
48 888-362-2447 | 216-765-5000 | usb.affymetrix.com
78355 50 RT rctns and 100 PCR rctns
The Two-Step RT-PCR Kit is designed for flexible RT-PCR in one- or two-tube formats.
�� Optimization of PCR independent of RT�� Analysis of the expression of multiple genes in individual RNA samples with oligo dT primers
Reverse transcription-polymerase chain reaction, or RT-PCR, is a method for converting and amplifying a single-stranded RNA template to yield abundant double-stranded complementary DNA (cDNA) product(1,2). In the RT step, the RT enzyme reverse transcribes an RNA template, yielding single-stranded cDNA. In the PCR step, a thermostable DNA polymerase amplifies the single-stranded cDNA to yield double-stranded cDNA product. RT-PCR may be carried out in several ways. One approach, often termed two-step RT-PCR, involves carrying out the RT (or cDNA synthesis) step in one tube and the PCR step in another tube(1) with gene specific primers, for detection of a single target from a single RNA sample. Another approach involves using one RT reaction, primed with oligo dT, for detection of multiple targets from the same RNA sample. A third approach, often termed one-step RT-PCR, involves setting up the RT and PCR simultaneously with gene specific primers and carrying out the reactions sequentially in one tube(2). This is particularly useful for detection of one or a few targets in multiple RNA samples. The Two-Step RT-PCR Kit has been designed for use in all of these types of RT-PCR.
All in one RT-PCR kitThe Two-Step RT-PCR Kit comes complete and ready-to-use for diverse RT-PCR applications. The kit can be used with RNA samples and custom primers. The kit uses M-MLV Reverse Transcriptase(3) and Taq DNA Polymerase(4), provided in individual tubes at concentrations optimized to balance sensitivity and specificity in two-step RT-PCR. Optimized RT and PCR reaction buffers, RNase Inhibitor, Ultrapure dNTPs, supplemental magnesium chloride, and RNase-free water are also included, allowing flex-ibility in setting up RT and PCR individually. A simple enzyme dilution step also allows use of the kit for one-step RT-PCR.
Highly sensitive, highly specificThe kit can be used for detection of diverse RNA targets based on generation of long cDNAs (to at least 5.6 kb) followed by amplification of short (~0.2 to 1.5 kb) PCR products (Fig. 1). Moderately and weakly expressed targets can generally be detected in 1 ng to 1 µg total RNA or 100 pg to 100 ng polyA RNA. Highly expressed targets can be detected in even lower amounts of RNA. Standard reaction conditions are sufficient for specific amplification of most targets. A few simple protocol adjustments, such as optimization of the amount of primers or addition of supplements for amplifying G+C rich targets, enable specific amplification for others(5).
Flexible formatA wide range of RT-PCR experiments are accessible due to the all-in-one nature of this kit. One set of reagents can be applied easily in multiple types of RT-PCR, allowing great flexibility in planning, carry-ing out, and optimizing experiments.
One kit, many applicationsThe Two-Step RT-PCR Kit is well suited for diverse RT-PCR applications. The two-step/gene specific primer format is excellent for qualitative analysis of gene expression, highly detailed analysis of RNA splice variants, and optimization of PCR indepen-dent of RT. The two-step/oligo dT primer format is excellent for analysis of the expression of multiple genes in individual RNA samples. And the one-step format is excellent for rapid analysis of single genes in multiple RNA samples. Depending on the scale of experiments and the size of PCR products, use of the One-Step RT-PCR Kit (PN 78350) coupled with the Taq PCR Master Mix or Taq PCR Kit may be more efficient. The Two-Step RT-PCR Kit (PN 78355) is one kit with many applications.
Kit components:M-MLV Reverse TranscriptaseTaq DNA PolymeraseRT Reaction Buffer (5X) (includes MgCl2)PCR Reaction Buffer (10X) (includes MgCl2)PCR Nucleotide Mix, Ultrapure: 10 mM each dATP,
dCTP, dGTP, dTTPRNase Inhibitor, Recombinant (4 units/µl)Magnesium Chloride (25 mM)RNase-Free (DEPC-treated) Water
Shipping and storage:Shipped on dry ice. Store at -20°C.
References:1. Sambrook, J. and Russel, D. W. (2001) “Molecular
Cloning: A Laboratory Manual,” Cold Spring Harbor Laboratory Press, 8.46-8.53.
2. Sellner, L. N., Coelen, R. J., and Mackenzie, J. S. (1992) Nucl. Acids Res. 20, 1487-1490.
3. Roth, M. J., Tanese, N., and Goff, S. P. (1985) J. Biol. Chem. 260, 9326-9335.
4. Saiki, R. K., Gelfand, D. H., Stoffel, S., Scharf, S. J., Higuchi, R., Horn, G. T., Mullis, K. B., and Erlich, H. A. (1988) Science 239, 487-491.
5. Tech Tip 206, Simple Approaches for Optimization of RT-PCR, USB Corporation, Cleveland, Ohio.
Related products
RT-PCR Master Mix (2X)78370 100 reactions
FideliTaq RT-PCR Master Mix (2X)71185 100 reactions
Two-Step RT-PCR Kit
Fig. 1. Amplification of diverse RNA targets by two-step RT-PCR. Target (source): β-actin (human liver), Numb (human liver), Ubiquitin (Arabidopsis leaf), and Terminal Deoxynucleotidyl Transferase (TdT) (calf thymus). RT was carried out on 1 µg total RNA with priming by oligo dT, and PCR was conducted on 10-1 dilution of RT reaction. Gene specific primers were designed to generate prod-ucts of particular sizes. M: Marker; Lanes 1-3: β-actin, 0.5 kb, 1.0 kb, 1.5 kb; Lanes 4-5: Numb 0.5 kb, 1.0 kb; Lane 6: Ubiquitin, 1.5 kb; Lane 7: TdT, 1.5 kb; Lane 8: Clathrin, 5.6 kb.
PCR Tools | Reverse Transcription RT-PCR—Kits
49For bulk or alternate pack sizes, email us at [email protected].
78250 20 reactions78200 100 reactions78201 500 reactions78202 2,000 reactions78205 5,000 reactionsCustom formulations by request
�� Conserves PCR samples – 100% recovery of both short and long PCR products�� One tube/one-step PCR cleanup – Add ExoSAP-IT reagent directly to PCR product.�� Eliminates spin columns – Decreases time and expense while increasing yield�� Removes contaminating primers and dNTPs – No interference in downstream applications�� Scalable – Economical for high-throughput purification�� Simple processing – Robotic-friendly; Replaces beads, filtrations, and plates�� Generates less waste than columns
ExoSAP-IT reagent is designed for simple, quick PCR cleanup for downstream applications, such as DNA sequencing or Single Nucleotide Polymorphism (SNP) analysis. When PCR amplification is complete, any unused dNTPs and primers remaining in the PCR product mixture will interfere with these methods. ExoSAP-IT PCR Cleanup removes these contaminants.
ExoSAP-IT reagent is added directly to the PCR product and incubated at 37°C for 15 minutes. After PCR treatment, it is inactivated simply by heating to 80°C for 15 minutes (Fig. 1).
ExoSAP-IT single-step PCR cleanup utilizes two hydrolytic enzymes, Exonuclease I and Shrimp Alkaline Phosphatase (SAP), together in a specially formulated buffer, to remove unwanted dNTPs and primers from PCR products. Exonuclease I removes residual single-stranded primers and any extraneous single-stranded DNA produced in the PCR. SAP removes the remaining dNTPs from the PCR mixture.
Rapid PCR product cleanup protocolThe ExoSAP-IT method requires only one pipet-ting step and two incubations. Just add ExoSAP-IT reagent to the PCR product and within 30 minutes sequencing or SNP analysis can be performed.
Simple: single-stepThe method is designed to require a minimum of “hands-on” time. Enzymatic removal of excess primers and unincorporated nucleotides occurs in one easy step by using ExoSAP-IT reagent in a single tube or microtiter well. Only simple pipette transfers are required, therefore, many samples can be pro-cessed at once, either manually or with robotics.
No sample lossUse of ExoSAP-IT reagent eliminates all gel or column purifications, sedimentations, filtrations, beads, and/or magnetic separations(1). There is 100% recovery of both short and long PCR products with ExoSAP-IT (Fig. 2).
Achieve high quality data from PCR productsExoSAP-IT reagent may be used as an effective cleanup method prior to fluorescent or radioactive DNA sequencing (Fig. 3), SNP analysis, or any other application requiring a PCR product free of excess nucleotides and primers.
Shipping and storage:Shipped on dry ice. Store at -20°C.
References:1. Dugan, K. A., Lawrence, H. S., Hares, D. R.,
Fisher, C. L. and Budowle B. (2002) J. Forensic Sci 47, 811-818.
2. Hanke, M. and Wink, M. (1994) BioTechniques 17, 858-860.
3. Mu, J., Duan, J., Makova, K., Joy, D., Huynh, C., Branch, O., Li, W. and Su, X. (2002) Nature 418, 323-326.
4. Silva, Jr., W. A., Costa, M. C. R., Valente, V., De Freitas Sousa, J., Pacó-Larson, M. L., Espreafico, E. M., Camargo, S. S., Monteiro, E., De Jesus, A., Holanda, M. A., Zago, M. A., Simpson, A. J. G. and Neto, E. D. (2001) BioTechniques 30, 537-542.
5. Werle, E., Scneider C., Renner, M., Völker, M. and Fiehn, W. (1994) Nucl. Acids Res. 22, 4354-4355.
ExoSAP-IT PCR Product Cleanup
Fig. 2. ExoSAP-IT treatment of PCR products with no sample loss. Single-copy targets were amplified from human genomic DNA. HES-1 (125 bp), numb (455 bp), NRAGE (1.55 kb), and numb (4.6 kb) were loaded on a 1.5% agarose gel before (pre) and after (post) ExoSAP-IT treatment. M is the DNA marker lane. Note that a variety of PCR product sizes can be treated with ExoSAP-IT, even a 125 bp fragment, with no sample loss.
HES-1 numb NRAGE numb M pre post pre post pre post pre post
12 kb –
2 kb –
1 kb –
100 bp –
PCR Mixture Post-Amplification
PCR Product
+
Nucleosides
+
Inorganic Phosphate (Pi)
Add ExoSAP-IT®
Excess Primers
Excess dNTPs
37°C, 15 min for treatment80°C, 15 min to inactivate
Fig. 1. Summary of ExoSAP-IT PCR product treatment.
Fig. 3. Fluorescent sequencing results of a 100 bp pUC18 PCR fragment sequenced with a -20 Fwd primer using fluo-rescent sequencing reagents. PCR cleanup performed with: (a) ExoSAP-IT; (b) a column designed for PCR cleanup. Base miscalls in (b) are due to inherently low yields of short PCR products when using columns.
a
b
PCR Tools | PCR Purification
Related product
HT ExoSAP-IT High-Throughput PCR Product Cleanup78395 480 rctns x 8-tube strip 5,760 rctns x 1 plate (12 x 8-tube strips) 23,040 rctns - 4 plates 1,000 rctns (2 ml) 5,000 rctns (10 ml)
50 888-362-2447 | 216-765-5000 | usb.affymetrix.com
78395 480 rctns x 8-tube strip 5,760 rctns x 1 plate (12 x 8-tube strips) 23,040 rctns - 4 plates 1,000 rctns (2 ml) 5,000 rctns (10 ml)
�� One-tube PCR cleanup – Add HT ExoSAP-IT reagent directly to PCR product.�� Exceptional accuracy – Achieve high quality data even with long read lengths (Fig. 1)�� 100% sample recovery – No loss of PCR products regardless of the fragment size (Fig. 2)�� Simple, single-step – Replace multiple steps and wasted time with bead or column use�� High-throughput processing – Even faster time to results with a low viscosity formulation, allowing robotic pipetting�� Removes excess primers and dNTPs – Does not interfere with downstream applications • sequencing • SNP analysis • single base extension • fragment analysis • in vitro transcription�� Scalable – Treat reaction volumes from 5 µl to 5 L�� Convenient packaging – Available in 8-tube strips and 12 x 8-tube strips in a 96-well plate �� Stable at +25°C for 8 hours – Retains full functional activity and at 4°C is stable for one week (Fig. 3)
HT ExoSAP-IT High-Throughput PCR Product Cleanup is an alternative formulation of ExoSAP-IT reagent specifically designed for the unique require-ments of high-throughput, automated platform and multi-channel pipettes. HT ExoSAP-IT reagent has both a longer lifetime at higher temperatures and a decreased viscosity for robotic pipetting. Like ExoSAP-IT PCR Product Cleanup (PN 78200), HT ExoSAP-IT reagent is a mixture of Exonuclease I and Shrimp Alkaline Phosphatase (SAP) which
removes excess primers and dNTPs following a PCR reaction in a single incubation. HT ExoSAP-IT reagent possesses greater thermal stability and lower viscosity which provides greater ease-of-use in automated 96- and 384-well formats.
HT ExoSAP-IT reagent is added directly to the PCR product and incubated at 37°C for 15 minutes. After PCR treatment, it is inactivated simply by heating to 80°C for 15 minutes. Our high-throughput formula-tion is designed for robotic pipetting and multi-channel pipettes. HT ExoSAP-IT reagent has a lower viscosity than the standard mix and is available in an 8-tube strip format for ease of use.
HT ExoSAP-IT reagent utilizes two hydrolytic enzymes, Exonuclease I and SAP, together in a specially formulated buffer, to remove unwanted dNTPs and primers from PCR products. Exonuclease I removes residual single-stranded primers and any extraneous single-stranded DNA produced in the PCR. SAP removes the remaining dNTPs from the PCR mixture.
Simple: Single-stepHT ExoSAP-IT reagent requires only one pipetting step and two incubations. Just add HT ExoSAP-IT reagent to the PCR product and within 30 minutes sequencing, SNP analysis, single base extension, or fragment analysis can be performed.
Enzymatic removal of excess primers and unincorpo-rated nucleotides occurs in one easy step by using HT ExoSAP-IT reagent in a single-tube, 8-tube strip, or 12 strips in a 96-well low skirted PCR plate. Our new formatting makes this an ideal product for use with robotics.
Exceptional accuracy with HT ExoSAP-IT Achieve high data quality and sequencing accuracy with HT ExoSAP-IT reagent, even with long read lengths. Sequence reads of PCR products treated with HT ExoSAP-IT reagent were on average 50+ bases longer than samples treated with competitor products. Using human genomic DNA, a 1,007 bp fragment was sequenced with no miscalls (Fig. 1). The size limitations associated with alternative PCR cleanup methods are not a factor with HT ExoSAP-IT reagent. Phred 20 values over the entire length of a 970 bp sequence was 822 ± 9 with HT ExoSAP-IT-treated samples and only 776 ± 83 with samples treated with Agencourt® AMPure® XP (see Table 1).
No sample lossUse of HT ExoSAP-IT eliminates all gel or column purifications, sedimentations, filtrations, beads, and/or magnetic separations(1). There is 100% recovery of both short and long PCR products with HT ExoSAP-IT High-Throughput PCR Product Cleanup (Fig. 2).
HT ExoSAP-IT High-Throughput PCR Product Cleanup
HT ExoSAP-IT
AMPure XP
3 miscalls
Fig. 2. Greater recovery of PCR product with HT ExoSAP-IT reagent. Equivalent volumes of PCR product were visualized on an ethid-ium bromide agarose gel and the band volume determined using image analysis. HT-ExoSAP-IT reagent allowed recovery of 100% of the 86 and 103 bp PCR products but AMPure XP beads allowed only 8 and 14% recovery, respectively. Un – Untreated, HT – HT-ExoSAP-IT treated, XP – AMPure XP purified, 100 bp – Affymetrix 100 bp ladder, bands from 100 to 1,000 bp by 100 bp intervals (PN 76712).
86 bp 103 bp100 bp Un HT XP Un HT XP
545 bp 1,007 bp100 bp Un HT XP Un HT XP
PCR Tools | PCR Purification
Fig. 1. Sequencing of a 1,007 bp treated PCR product. A 1,007 bp fragment was amplified and treated with HT ExoSAP-IT reagent (above) or Agencourt AMPure XP beads (below) and sequenced. Pherograms revealed no miscalls with HT ExoSAP-IT but three miscalls with Agencourt AMPure XP beads at position 203, 204, and 220.
Phred score values of samples processed with HT ExoSAP-IT and AMPure XP Beads
Table 1. HT ExoSAP-IT reagent gives better sequencing results.
HT ExoSAP-ITAMPure XP beads
Number bases read (Average of 16 samples)
901 850
Total miscalls 0 25
Pass rate (%)* 100% 99.7%
Phred 20** 822 ± 9 776 ± 83
* Pass rate – Average Phred value greater than 20 bases between 100 bp and 300 bp.** Phred 20 values over the entire length of sequence (no trimming ~970 bp)
51For bulk or alternate pack sizes, email us at [email protected].
HT ExoSAP-IT-treated PCR product is stableHT ExoSAP-IT reagent is rigorously tested and subject to strict quality control. Fig. 3 demonstrates that no product degradation occurred after storage of HT ExoSAP-IT-treated PCR product for one week at 25°C.
Storage:Shipped on dry ice. Store at -20°C.
References:1. Dugan, K. A., Lawrence, H. S., Hares, D. R.,
Fisher, C. L. and Budowle B. (2002) J. Forensic Sci 47, 811-818.
2. Hanke, M. and Wink, M. (1994) BioTechniques 17, 858-860.
3. Mu, J., Duan, J., Makova, K., Joy, D., Huynh, C., Branch, O., Li, W. and Su, X. (2002) Nature 418, 323-326.
4. Silva, Jr., W. A., Costa, M. C. R., Valente, V., De Freitas Sousa, J., Pacó-Larson, M. L., Espreafico, E. M., Camargo, S. S., Monteiro, E., De Jesus, A., Holanda, M. A., Zago, M. A., Simpson, A. J. G. and Neto, E. D. (2001) BioTechniques 30, 537-542.
5. Werle, E., Scneider C., Renner, M., Völker, M. and Fiehn, W. (1994) Nucl. Acids Res. 22, 4354-4355.
Related products
ExoSAP-IT PCR Product Cleanup78250 20 reactions78200 100 reactions78201 500 reactions78202 2,000 reactions78205 5,000 reactions
VeriQuest Fast Probe qPCR Master Mix (2X)75680 100 reactions 500 reactions 1,000 reactions 2,500 reactions 5,000 reactions
VeriQuest Fast Probe qPCR Master Mix, No Reference Dye (2X)75685 100 reactions 500 reactions 1,000 reactions 2,500 reactions 5,000 reactions
VeriQuest Fast SYBR Green qPCR Master Mix (2X)75690 100 reactions 500 reactions 1,000 reactions 2,500 reactions 5,000 reactions
VeriQuest Fast SYBR Green qPCR Master Mix with Fluorescein (2X)75675 100 reactions 500 reactions 1,000 reactions 2,500 reactions 5,000 reactions
VeriQuest Probe qPCR Master Mix (2X)75650 40 reactions 200 reactions 400 reactions 1,000 reactions 2,000 reactions
VeriQuest Probe qPCR Master Mix, No Reference Dye (2X)75660 40 reactions 200 reactions 400 reactions 1,000 reactions 2,000 reactions
VeriQuest SYBR Green qPCR Master Mix (2X)75600 40 reactions 200 reactions 400 reactions 1,000 reactions 2,000 reactions
VeriQuest SYBR Green qPCR Master Mix with Fluorescein (2X)75665 40 reactions 200 reactions 400 reactions 1,000 reactions 2,000 reactions
VeriQuest Taq DNA Polymerase71170 50 units 250 units 1,000 units 5,000 units
FideliTaq DNA Polymerase71180 50 units 250 units 1,000 units 5 x 250 units 5,000 units
RubyTaq DNA Polymerase71190 50 units 250 units 1,000 units 5,000 units
PCR Nucleotide Mix, 10 mM Solution(10 mM each dATP, dCTP, dGTP, dTTP)77212 500 µl
Fig. 3. High stability of HT ExoSAP-IT-treated products. A 1,007 bp DNA fragment amplified from genomic DNA with HotStart FideliTaq Master Mix (PN 71156) was treated with HT ExoSAP-IT reagent and stored for seven days at: -20ºC (lane 1), 4ºC (lane 2), or 25ºC (lane 3). Samples were visualized on 1.5% agarose/TAE/ethid-ium bromide gel. Lane MW: DNA Ladder, 1 kb Plus (PN 76714).
1 kb -20°C 4°C 25°C
HT ExoSAP-IT High-Throughput PCR Product Cleanup continued...
PCR Tools | PCR Purification
52 888-362-2447 | 216-765-5000 | usb.affymetrix.com
70995 100 reactions70996 500 reactions70997 2,000 reactionsCustoms by request
The PCR Product Pre-Sequencing Kit uses a novel enzymatic cleanup method to pre-treat PCR products prior to sequencing without any subsequent purifica-tion or separation steps. The two hydrolytic enzymes used in this kit, Shrimp Alkaline Phosphatase (SAP) and Exonuclease I, effectively remove excess dNTPs and primers present in the final PCR product reac-tion mixture. The enzymes are conveniently added to an aliquot of the PCR product mixture, incubated at 37°C, and then inactivated at 80°C. The result is a cleaner PCR product, free from excessive nucleotides and primers, ready to be sequenced with standard sequencing reagents.
Achieve high sequence quality with PCR productsEnzymatic treatment to remove excess primers and nucleotides yields templates which can be easily sequenced. Problems with left-over PCR primers leading to background bands are virtually elimi-nated. The kit may be used as an effective cleanup method prior to any DNA sequencing protocol, including but not limited to the Sequenase Version 2.0 DNA Sequencing Kit (PN 70770) or the Thermo Sequenase Cycle Sequencing Kits (PNs 78500 and 79260).
Kit components:Shrimp Alkaline PhosphataseExonuclease I
Shipping and storage:Shipped on dry ice. Store at -20°C.
References:1. Hanke, M. and Wink, M. (1994) BioTechniques
17, 858-860.2. Werle, E., Scneider C., Renner, M., Völker, M. and
Fiehn, W. (1994) Nucl. Acids Res. 22, 4354-4355.
Related products
Sequenase™ Version 2.0 DNASequencing Kit70770 100 reactions
Thermo Sequenase™ Cycle Sequencing Kit78500 100 reactions
PCR Product Pre-Sequencing Kit
78260 864 µl
SBE Cleanup Reagent is a specialized formulation of ExoSAP-IT PCR Product Cleanup (PN 78200), which is based on enzymatic degradation of unconsumed primers and dNTPs present in PCR products. SBE Cleanup Reagent has been designed for use with the Beckman Coulter BioSciences SNPware™ Core Reagent Kit (PN 101-04-300).
Single-stepThe SBE Cleanup Reagent is designed for simple, quick PCR cleanup. SBE Cleanup Reagent utilizes two hydrolytic enzymes, Exonuclease I and Shrimp Alkaline Phosphatase (SAP), together in a specially formulated buffer, to remove unwanted dNTPs and primers from PCR products. Exonuclease I removes residual single-stranded primers and any extraneous
single-stranded DNA produced in the PCR. SAP removes the remaining dNTPs from the PCR mixture. It is added directly to the PCR product. SBE Cleanup Reagent is active in the buffer used for PCR, so no buffer exchange is required.
Shipping and storage:Shipped on dry ice. Store at -20°C.
SBE Cleanup Reagent
Binding capacity up to 15 µg78758 50 preps78759 250 prepsSee page 146
Designed for the purification of DNA from reactions or for buffer exchange
�� Spin columns based on familiar silica-membrane technology�� High recovery rates up to 90% with low elution volume (15 μl)
�� Ideal for cleanup of reaction products�� Binding capacity up to 15 μg�� For DNA fragments 50 – 10,000 bp
The PrepEase DNA Cleanup Kit is designed to purify DNA fragments from enzymatic reactions or for buffer exchange. Thus, contaminants such as salts, enzymes, nucleotides, labeling reagents, etc. can be removed from a DNA sample. This kit can also be used for purification of DNA from Chromatin Immunoprecipitation (ChIP) assays.
Each kit includes PrepEase Cleanup Columns, collection tubes and the necessary reagents for the purification of DNA fragments.
PrepEase DNA Cleanup Kits
Binding capacity up to 15 µg78756 50 preps78757 250 prepsSee page 146
Designed for purifying DNA fragments from agarose gels. Also suited for PCR product purification.
�� Spin columns based on familiar silica-membrane technology�� High recovery rates of up to 90% with low elution volume (15 µl)�� Ideal for purifying restriction-digested DNA in gels or PCR products
The PrepEase Gel Extraction Kit may be used to purify DNA from agarose gels or PCR reaction mixtures. In addition, the kit may be used for desali-nation, and the removal of enzymes, nucleotides or labeling reagents from sample DNA.
PrepEase Gel Extraction Kits
PCR Tools | PCR Purification
53For bulk or alternate pack sizes, email us at [email protected].
78761 10 x 96-well plates78762 50 x 96-well platesSee page 148
Designed for high-throughput purification of PCR products using 96-well plates.
�� For PCR products ≥ 150 bp�� Process 20 - 300 μl PCR volume per well�� Recover up to 95% depending on length of PCR product in as little as 25 μl
�� May be performed manually by vacuum, by centrifugation or by using an automated workstation�� Includes 96-well plates and buffer for PCR product purification
PrepEase PCR Purification 96-Well Plate Kits (Ultrafiltration)
75767 500 μl
Fluorescein Passive Reference Dye is specially formu-lated for use on Bio-Rad real-time PCR instruments such as the MyiQ, iCycler iQ, and iQ5. This inert dye may be added to quantitative, real-time PCR reac-tions containing SYBR Green I. Fluorescein allows the collection of Dynamic Well Factors from the experimental plate, which is the preferred method. This normalizes the well-to-well differences that may occur due to artifacts such as pipetting errors or instrument limitations.
Fluorescein Passive Reference Dye is composed of a 500 nM solution of fluorescein in 10 mM Tris-HCl (pH 8.6), 0.1 mM EDTA, and 0.01% Tween®-20. Generally, the optimum final fluorescein concentra-tion is 10 nM but a titration between 5-20 nM may be necessary depending on the particular instru-ment, the amount of SYBR in the reaction mix, and the desired background. See specific instrument instructions for further details on well factor collection. The following table shows current recommendations from Affymetrix.
Spectral Characteristics:Excitation λ Maximum: ≈ 495 nmEmission λ Maximum: ≈ 515 nm
Shipping and storage:Shipped on dry ice. Store at -20°C. Protect from light.
Fluorescein Passive Reference Dye
Fluorescein Final Concentration Amount per 50 μl reaction Final Concentration Dilution Factor
1.0 μl (0.5 - 2.0 μl) 10 nM (5 - 20 nM) 50X
Subtract the volume of the dye from the volume of water needed to prepare the PCR reaction.
75768 500 μl
ROX Passive Reference Dye is specially formulated for use on Applied Biosystems (ABI) and Stratagene real-time PCR instruments. This inert dye, whose fluorescence does not change during the reaction, may be added to quantitative, real-time PCR reac-tions to normalize the well-to-well differences that may occur due to artifacts such as pipetting errors or instrument limitations.
ROX Passive Reference Dye is composed of a 25 μM solution of 5-carboxy-X-rhodamine in 10 mM Tris-HCl (pH 8.6), 0.1 mM EDTA, and 0.01% Tween-20.
The following table shows current recommendations from Affymetrix.
Spectral Characteristics:Excitation λ Maximum: ≈ 575 nmEmission λ Maximum: ≈ 600 nm
Shipping and storage:Shipped on dry ice. Store at -20°C. Protect from light.
ROX Passive Reference Dye
ROX Final Concentration for Different Instruments Amount per Final ROX Dilution Instrument 50 μl reaction Concentration Factor
ABI 7000, 7300, 7700, 1.0 μl 500 nM 50X 7900HT, 7900HT Fast, (0.6 - 1.0 μl) (300 - 500 nM) StepOne™ and StepOne Plus™
ABI 7500 and ABI 7500 Fast; 0.1 μl 50 nM 500X Stratagene Mx3000™, (0.06 - 0.1 μl)* (30 - 50 nM) Mx3005P™, and Mx4000™
Subtract the volume of the dye from the volume of water needed to prepare the PCR reaction.*Note: ROX may be diluted 1:10 in water to aid in accurate pipetting.
PCR Tools | PCR Purification/Passive Reference Dyes
NucleotidesDeoxynucleotides
PCR Nucleotide Mixes
Dideoxynucleotides
Ribonucleotides
Labeled Nucleotides
55For bulk or alternate pack sizes, email us at [email protected].
USB Ultrapure Nucleotide Solutions
�� Functionally tested: All USB Ultrapure dNTPs are functionally tested in long PCR to generate a 20.7 kb fragment.�� Convenient packaging: dNTPs are available separately, as a set of 4, or as a PCR mix.�� Bulk sizes and custom blends available upon request.
Deoxynucleotides (dNTPs), Sodium Salt Aqueous Solutions, pH 7.5
Prod. No. Product Description [conc] Size
77102 dATP 100 mM 25 µmol (250 µl) 2'-Deoxyadenosine-5'-Triphosphate 100 µmol (1 ml) Ultrapure 500 µmol (5 ml)
77104 dCTP 100 mM 25 µmol (250 µl) 2'-Deoxycytidine-5'-Triphosphate 100 µmol (1 ml) Ultrapure 500 µmol (5 ml)
77106 dGTP 100 mM 25 µmol (250 µl) 2'-Deoxyguanosine-5'-Triphosphate 100 µmol (1 ml) Ultrapure 500 µmol (5 ml)
77108 dTTP 100 mM 25 µmol (250 µl) 2'-Deoxythymidine-5'-Triphosphate 100 µmol (1 ml) Ultrapure 500 µmol (5 ml)
77206 dUTP 100 mM 25 µmol (250 µl) 2'-Deoxyuridine-5'-Triphosphate 100 µmol (1 ml) Ultrapure 500 µmol (5 ml)
77100 dNTPs, Sets of Four (dA, dC, dG, dT) 100 mM 4 x 25 µmol (250 µl) 2'-Deoxynucleoside-5'-Triphosphates, Ultrapure 4 dNTPs per pack (pk) 1 pk77328 100 mM 4 x 100 µmol (1 ml) 4 dNTPs per pack (pk) 1 pk77128 100 mM 4 x 500 µmol (5 ml) 4 dNTPs per pack (pk) 1 pk
77212 PCR Nucleotide Mix, 10 mM Solution 10 mM 500 µl Ultrapure 10 mM 2 x 500 µl 10 mM each dATP, dCTP, dGTP, dTTP
77119 PCR Nucleotide Mix, 25 mM Solution 25 mM 500 µl Ultrapure 25 mM each dATP, dCTP, dGTP, dTTP
77330 PCR Nucleotide Mix with dUTP, 10 mM Solution 10 mM 500 µl Ultrapure 10 mM each dATP, dCTP, dGTP, dUTP
Deoxynucleotides (dNTPs), Lithium Salt Aqueous Solutions, pH 7.0
Prod. No. Product Description [conc] Size
70065 7-deaza-dGTP 10 mM 2 µmol (200 µl) 7-deaza-2'-Deoxyguanosine-5'-Triphosphate Ultrapure
Nucleotides
56 888-362-2447 | 216-765-5000 | usb.affymetrix.com
Dideoxynucleotides (ddNTPs), Sodium Salt Aqueous Solutions, pH 7.5�� Functionally tested: USB ddNTPs are functionally tested to meet or exceed criteria for high quality sequencing.�� Bulk sizes and custom blends available upon request.
Prod. No. Product Description [conc] Size
77110 ddATP 10 mM 0.5 µmol (50 µl) 2',3'-Dideoxyadenosine-5'-Triphosphate 1 µmol (100 µl) Ultrapure
77112 ddCTP 10 mM 0.5 µmol (50 µl) 2',3'-Dideoxycytidine-5'-Triphosphate 1 µmol (100 µl) Ultrapure
77116 ddTTP 10 mM 0.5 µmol (50 µl) 2',3'-Dideoxythymidine-5'-Triphosphate 1 µmol (100 µl) Ultrapure
77126 ddNTPs, Set of Four (ddA, ddC, ddG, ddT) 10 mM 4 x 0.5 µmol (50 µl) Dideoxynucleoside Triphosphates 4 ddNTPs per pack (pk) Ultrapure 1 pk77125 10 mM 4 x 1 µmol (100 µl) 4 ddNTPs per pack (pk) 1 pk77335 100 mM 4 x 4 µmol (40 µl) 4 ddNTPs per pack (pk) 1 pk
Ribonucleotides (NTPs), Sodium Salt Aqueous Solutions, pH 7.5
Prod. No. Product Description [conc] Size
77241 ATP 100 mM 25 µmol (250 µl) Adenosine-5'-Triphosphate Ultrapure
77243 GTP 100 mM 25 µmol (250 µl) Guanosine-5'-Triphosphate Ultrapure
77245 NTPs, Set of Four (ATP, CTP, GTP, UTP) 100 mM 4 x 25 µmol (250 µl) Nucleoside Triphosphates 4 NTPs per pack (pk) Ultrapure 1 pk
USB Ultrapure Nucleotide Solutions
Nucleotides
57For bulk or alternate pack sizes, email us at [email protected].
Nucleotides | Nucleotide Labeling Reagents
Molecular λ max Absorbance at λ max Compound Weight (pH 7.0) 1 M solution (pH 7.0)
ATP 507.2 259 15,400
CTP 483.2 271 9,000
GTP 523.2 253 13,700
UTP 484.2 262 10,000
dATP 491.2 259 15,200
dCTP 467.2 271 9,300
dGTP 507.2 253 13,700
dTTP 482.2 267 9,600
Nucleotide Physical Properties
79015 250 nmol (10 mM) 1,000 nmol (10 mM)
DNA Labeling Reagent, DLR, is a proprietary biotin-labeled, abasic, deoxynucleotide analog designed for efficient 3’ end-labeling of DNA using Terminal Deoxynucleotidyl Transferase (TdT) after amplification reactions such as PCR. When used with streptadivin conjugated to a reporter, the 3’ biotin labeled DNA
fragment can be detected by enzyme activity, fluo-rescence, or by the binding of a secondary antibody. In addition, DLR will not base-pair with natural nucleotides, thus reducing non-specific binding and maximizing the signal-to-noise ratio. DLR is a com-ponent of the Affymetrix GeneChip DNA labeling reagent (PN 900542) and has been used extensively to successfully probe microarrays for decades.
C24H40Li4N5O16P3SFormula Weight: 807.35
Product specifications:Purity: >95%Concentration: 10 mMAppearance: Clear, colorless liquid
Storage: Store at -20°C, avoid freeze/thaw cycles
Biotin-11-dXTP Analog (DNA Labeling Reagent, DLR)
79010 250 nmol (10 mM) 1,000 nmol (10 mM)
RNA Labeling Reagent, RLR, is a proprietary biotin-labeled pseudo-uridine triphosphate designed for incorporation into an RNA strand during an in vitro transcription reaction. This creates an RNA target internally labeled with biotin that can be detected
using streptadivin conjugated to fluorophores, enzymes, or antibodies. RLR is designed for efficient incorporation by T7 RNA polymerase and has been extensively tested in the Affymetrix GeneChip IVT labeling kit (PN 900449). The reagent is optimized for use in high throughput applications and probing high density microarrays.
C30H44Li4N7O19P3SFormula Weight: 959.46
Product specifications:Purity: >95%Concentration: 10 mMAppearance: Clear, colorless liquid
Storage: Store at -20°C, avoid freeze/thaw cycles
Biotin-11-ψTP Analog (RNA Labeling Reagent, RLR)
58 888-362-2447 | 216-765-5000 | usb.affymetrix.com
Copyright 1983 from Molecular Biology of the Cell, by Alberts et al. Reproduced by permission of Garland Science/Taylor & Francis LLC.
Structures of Nucleotides and Nucleosides
Nucleotides
59For bulk or alternate pack sizes, email us at [email protected].
2'-Deoxyadenosine-5'-Triphosphate, Sodium Salt, 100 mM Solution (dATP)
ULTRAPUREC10H12N5O12P3Na4
Formula Weight: 579.11Product specifications:Form: 100 mM aqueous solutionAssay (HPLC): ≥ 99%pH: 7.5 ± 0.2λ max: 259 nm ± 1 nmSpectral Ratios: A250/A260: 0.80 ± 0.03 A280/A260: 0.12 ± 0.02Functional Tests: Functionally tested in long PCR to
generate a 20.7 kb fragment.Storage: -20°C
77102 25 µmol (250 µl) 100 µmol (1 ml) 500 µmol (5 ml)
2'-Deoxycytidine-5'-Triphosphate, Sodium Salt, 100 mM Solution (dCTP)
ULTRAPUREC9H12N3O13P3Na4
Formula Weight: 555.08Product specifications:Form: 100 mM aqueous solutionAssay (HPLC): ≥ 99%pH: 7.5 ± 0.2λ max (pH 2.0): 280 nm ± 1 nmSpectral Ratios (pH 2.0): A250/A260: 0.45 ± 0.03 A280/A260: 2.10 ± 0.15 A290/A260: 1.60 ± 0.10Functional Tests: Functionally tested in long PCR to
generate a 20.7 kb fragment.Storage: -20°C
77104 25 µmol (250 µl) 100 µmol (1 ml) 500 µmol (5 ml)
2'-Deoxyguanosine-5'-Triphosphate, Sodium Salt, 100 mM Solution (dGTP)
ULTRAPUREC10H12N5O13P3Na4
Formula Weight: 595.11Product specifications:Form: 100 mM aqueous solutionAssay (HPLC): ≥ 99%pH: 7.5 ± 0.2λmax: 252 nm ± 1 nmSpectral Ratios (pH 7.5): A250/A260: 1.18 ± 0.04 A280/A260: 0.67 ± 0.03 A290/A260: 0.28 ± 0.03Functional Tests: Functionally tested in long PCR to
generate a 20.7 kb fragment.Storage: -20°C
77106 25 µmol (250 µl) 100 µmol (1 ml) 500 µmol (5 ml)
2'-Deoxythymidine-5'-Triphosphate, Sodium Salt, 100 mM Solution (dTTP)
ULTRAPUREC10H13N2O14P3Na4
Formula Weight: 570.10Product specifications:Form: 100 mM aqueous solutionAssay (HPLC): ≥ 99%pH: 7.5 ± 0.2λmax: 267 nm ± 1 nmSpectral Ratios (pH 7.5): A250/A260: 0.65 ± 0.03 A280/A260: 0.73 ± 0.03 A290/A260: 0.28 ± 0.03Functional Tests: Functionally tested in long PCR to
generate a 20.7 kb fragment.Storage: -20°C
77108 25 µmol (250 µl) 100 µmol (1 ml) 500 µmol (5 ml)
2'-Deoxyuridine-5'-Triphosphate, Sodium Salt, 100 mM Solution (dUTP)
ULTRAPUREC9H11N5O12P3Na4
Formula Weight: 556.1Product specifications:Form: 100 mM aqueous solutionpH: 7.5 ± 0.2Assay (HPLC): ≥ 99%λmax: (pH 7.5) 262 nm ± 1 nmAbsorbance Ratios: A250/A260: 0.74 ± 0.03 A280/A260: 0.35 ± 0.03Functional Test: Functionally tested in PCRStorage: -20°C
77206 25 µmol (250 µl) 100 µmol (1 ml) 500 µmol (5 ml)
2'-Deoxynucleoside-5'-Triphosphates, Sodium Salt, 100 mM Solutions, Sets of Four (dNTPs)
ULTRAPUREForm: 100 mM aqueous solutions, pH 7.5; dATP,
dCTP, dGTP, dTTPFunctional Tests: Functionally tested in long PCR to
generate a 20.7 kb fragment.Storage: -20°C
4 x 25 µmol (250 µl) each dNTP per pack77100 1 pk4 x 100 µmol (1 ml) each dNTP per pack77328 1 pk4 x 500 µmol (5 ml) each dNTP per pack77128 1 pk
For powder forms of the above nucleotides and our complete selection of nucleotides, nucleosides, purines and pyrimidines, please see the Biochemicals section of this catalog.
Nucleotides | Deoxynucleotides (dNTPs)
60 888-362-2447 | 216-765-5000 | usb.affymetrix.com
PCR Nucleotide Mix, 10 mM SolutionULTRAPURE
Form: Premixed dNTPs at 10 mM each dATP, dCTP, dGTP, dTTP.
pH: 7.5Functionally tested in PCR with USB Taq DNA
Polymerase (PN 71160).Storage: -20°C
77212 500 µl 2 x 500 µl
PCR Nucleotide Mix, 25 mM SolutionULTRAPURE
Form: Premixed dNTPs at 25 mM each dATP, dCTP, dGTP, dTTP.
pH: 7.5Functionally tested in PCR with USB Taq DNA
Polymerase (PN 71160).Storage: -20°C
77119 500 µl
PCR Nucleotide Mix with dUTP, 10 mM SolutionULTRAPURE
Form: Premixed dNTPs at 10 mM each dATP, dCTP, dGTP, dUTP
pH: 7.5Functionally tested in PCR with USB Taq DNA
Polymerase (PN 71160).Storage: -20°C
77330 500 µl
7-deaza-2'-Deoxyguanosine-5'-Triphosphate, Lithium Salt, 10 mM Solution (7-deaza-dGTP)
ULTRAPUREC11H15N4O13P3Li2Formula Weight: 518.10Product specifications:Form: 10 mM aqueous solutionAssay (HPLC): ≥ 95%pH: 7.0λmax (pH: 7.5): 259 nm ± 1 nmFunctional Tests: Meets or exceeds criteria for high
quality sequencing.Storage: -20°C
70065 2 µmol
2',3'-Dideoxyadenosine-5'-Triphosphate (ddATP)
ULTRAPUREC10H12N5O11P3Na4
Formula Weight: 563.11Product specifications:Assay (FPLC): ≥ 99%pH: 7.5 ± 0.2λmax: 259 nm ± 1 nmSpectral Ratios (pH 7.5): A250/A260: 0.80 ± 0.03 A280/A260: 0.12 ± 0.02Functional Test: Meets or exceeds criteria for high
quality sequencing.Storage: -20°C
10 mM aqueous solution, pH 7.577110 0.5 µmol (50 µl) 1 µmol (100 µl)
2',3'-Dideoxycytidine-5'-Triphosphate (ddCTP)
ULTRAPUREC9H12N3O12P3Na4
Formula Weight: 539.08Product specifications:Assay (FPLC): ≥ 99%pH: 7.5 ± 0.2λmax (pH 2.0): 280 nm ± 1 nmSpectral Ratios (pH 2.0): A250/A260: 0.45 ± 0.03 A280/A260: 2.20 ± 0.20 A290/A260: 1.70 ± 0.20Functional Test: Meets or exceeds criteria for high
quality sequencing.Storage: -20°C
10 mM aqueous solution, pH 7.577112 0.5 µmol (50 µl) 1 µmol (100 µl)
2',3'-Dideoxythymidine-5'-Triphosphate (ddTTP)
ULTRAPUREC10H13N2O13P3Na4
Formula Weight: 554.10Product specifications:Assay (FPLC): ≥ 99%pH: 7.5 ± 0.2λmax (pH 7.5): 267 nm ± 1 nmSpectral Ratios (pH 7.5): A250/A260: 0.65 ± 0.03 A280/A260: 0.83 ± 0.03Functional Tests: Meets or exceeds criteria for high
quality sequencing.Storage: -20°C
10 mM aqueous solution, pH 7.577116 0.5 µmol (50 µl) 1 µmol (100 µl)
2',3'-Dideoxynucleoside-5'-Triphosphates, 10 mM, Sets of Four (ddNTPs)
ULTRAPUREForm: 10 mM aqueous solutions, pH 7.5; ddATP,
ddCTP, ddGTP, ddTTPFunctional Tests: Meets or exceeds criteria for high
quality sequencing.Storage: -20°C
4 x 0.5 µmol (50 µl) each dNTP per pack77126 1 pk4 x 1.0 µmol (100 µl) each dNTP per pack77125 1 pk
2',3'-Dideoxynucleoside-5'-Triphosphates, 100 mM, Set of Four (ddNTPs)
ULTRAPUREForm: 100 mM aqueous solutions, pH 7.5; ddATP,
ddCTP, ddGTP, ddTTPFunctional Tests: Meets or exceeds criteria for high
quality sequencing.Storage: -20°C
4 x 4 µmol (40 µl) each ddNTP per pack77335 1 pk
Custom blends available upon request
For powder forms of the above nucleotides and our complete selection of nucleotides, nucleosides, purines and pyrimidines, please see the Biochemicals section of this catalog.
Nucleotides | Deoxynucleotides (dNTPs) & Dideoxynucleotides (ddNTPs)
61For bulk or alternate pack sizes, email us at [email protected].
Adenosine-5'-Triphosphate, 100 mM Solution (ATP)
ULTRAPUREC10H12N5O13P3Na4
Formula Weight: 595.11Form: 100 mM aqueous solution, pH 7.5Storage: -20°C
77241 25 µmol (250 µl)
Guanosine-5'-Triphosphate, Sodium Salt, 100 mM Solution (GTP)
ULTRAPUREC10H12N5O14P3Na4
Formula Weight: 611.11Form: 100 mM aqueous solution, pH 7.5Storage: -20°C
77243 25 µmol (250 µl)
Nucleoside-5'-Triphosphates (ATP, CTP, GTP, UTP), Set of Four, 100 mM Solutions (NTPs)
ULTRAPURE100 mM aqueous solutions, pH 7.5Storage: -20°C
4 x 25 µmol (250 µl) each NTP per pack.77245 1 pk
Related products – RNA analysis
Oligo(dT)12-18 Primer, MW ≅450077405 100 µl
Oligo p(dT)12-18, Sodium Salt19817 5 units 25 units 100 units
Poly(A) Polymerase, Yeast74225Y 10,000 units74225Z 25,000 units
T3 RNA Polymerase70051Y 1,000 units70051Z 5,000 units
T7 RNA PolymeraseStandard Concentration, 20 units/µl70047Y 6,000 unitsHigh Concentration, >200 units/µl70001Z 30,000 units
Nucleotides | Ribonucleotides (NTPs)
For powder forms of the above nucleotides and our complete selection of nucleotides, nucleosides, purines and pyrimidines, please see the Biochemicals section of this catalog.
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Molecular Biology EnzymesBinding Proteins
Glycosylases
Kinases
Ligases
Nucleases
DNases
Endonucleases
Exonucleases
RNases
Phosphatases
Alkaline Phosphatases
Pyrophosphatase
Polymerases
DNA Polymerases
Reverse Transcriptases
RNA Polymerases
Transferase
RNase Inhibitors
Topoisomerase
63For bulk or alternate pack sizes, email us at [email protected].
Molecular Biology Enzymes | Table of Contents
Enzyme Applications 64
Binding Proteins RecAProtein 65Single-StrandedDNABindingProtein(SSB) 65T4Gene32Protein 66
GlycosylasesUracil-DNAGlycosylase,E. coli 67Uracil-DNAGlycosylase,Heat-Labile 67
KinasesOptiKinase™ 68T4PolynucleotideKinase(PNK) 69
LigasesE. coliDNALigase 70T4DNALigase 70T4RNALigase 71T4RNALigase2 71
Nucleases - DNasesDNaseI,Lyophilized 72DNaseI,Solution 72rDNaseI,RNase-Free 73ShrimpDNase,Recombinant 7410XDNaseIBuffer 74
Nucleases - EndonucleasesHumanApurinic/ApyrimidinicEndonuclease1
(APE1) 75T4EndonucleaseVII 75
Nucleases - ExonucleasesExonucleaseI 76ExonucleaseIII 77ExonucleaseVII 77T7Gene6Exonuclease 78
Nucleases - RNasesRNaseI 79RNaseI‘A’,Powder 79RNaseA,Lyophilized 79RNaseA,MBGrade,Solution 80RNaseH 80
Nucleases - Non-SpecificMicrococcalNuclease 81PhosphodiesteraseI 81
Phosphatases - Alkaline PhosphatasesCalfIntestinalAlkalinePhosphatase(CIAP) 82ShrimpAlkalinePhosphatase(SAP) 83SuperSAP™ShrimpAlkalinePhosphatase 84
Phosphatases - PyrophosphatasePyrophosphatase,Inorganic(Recombinant)
(rPPase) 85
Polymerases - DNA PolymerasesExonuclease-FreeKlenow 86Exonuclease-FreeKlenowTrisBuffer 86DNAPolymeraseI 87KlenowDNAPolymeraseI 87Sequenase™Version2.0DNAPolymerase 88T7DNAPolymerase 88TthDNAPolymerase 89
Polymerases - Reverse TranscriptasesAMVReverseTranscriptase 90M-MLVReverseTranscriptase 90
Polymerases - RNA PolymerasesE. coliRNAPolymeraseHoloenzyme 91Poly(A)Polymerase,Yeast 91T3RNAPolymerase 92T7RNAPolymerase 92
Polymerases - TransferaseTerminalDeoxynucleotidylTransferase,
Recombinant(rTdT) 93
RNase InhibitorsRNaseInhibitor(HumanPlacenta) 94RNaseInhibitor(Recombinant) 94
TopoisomeraseTopoisomeraseII,Alpha 95
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Molecular Biology Enzymes | Enzyme Applications
Application Enzyme
Blunt-endgeneration byremovalof3'protrudingends................T4andT7DNAPolymerase,Klenowfragment*,Blunt-ITRepairKit* byfillingin3'recessedends ...................T4andT7DNAPolymerase;Klenowfragment,Blunt-ITRepairKit*cDNAsynthesis fromRNA .................................ReverseTranscriptase;AMVRTorM-MLVRT,TthDNAPolymerase* ofsecondstrand ............................Klenowfragment,Sequenase,TaqDNAPolymerase,FideliTaqDNAPolymerase*CloningofDNAfragments .........................ShrimpAlkalinePhosphatase(SAP)*;CIP;T4DNALigase;Klenowfragment;restrictionendonucleasesDegradationofdouble-&single-strandedDNA*.........DNaseI*Degradationofdouble-strandedDNA,leaving...........ShrimpDNase* single-strandedDNAintact*DegradationofDNA Exonucleases—double-stranded 5'→3'....................................T7Gene6andLambdaExonuclease 3'→5'....................................ExonucleaseIII 5'→3'and3'→5'...........................Bal 31Nuclease,ExonucleaseV* Exonucleases—single-stranded 3'→5'....................................ExonucleaseI 5'→3'and3'→5'...........................ExonucleaseVII*,ExonucleaseV*DegradationofRNA..............................RibonucleaseAand/orT1;micrococcalnucleaseDegradationofRNAinRNA:DNAduplexes.............RibonucleaseHEndonucleases—double-stranded non-specific................................DNaseI,Bal31,andmicrococcalnucleases specific ...................................RestrictionendonucleasesEndonucleases—single-stranded ....................S1,Bal 31andmungbeannucleasesDephosphorylation...............................ShrimpAlkalinePhosphatase(SAP)*,BAPandCIPDNAsequencing.................................ModifiedT7DNAPolymerase(Sequenase*);TaqPolymerase;Klenowfragment;T4PolynucleotideKinase;
ReverseTranscriptase;ThermoSequenase*Intronmapping..................................ExonucleaseVII;S1NucleaseLabelingDNAat5'ends...........................T4PolynucleotideKinase,OptiKinase*LabelingDNAat3'ends...........................ModifiedT7DNAPolymerase(Sequenase*);Klenowfragment;ReverseTranscriptase;TerminalTransferaseLabelingRNAat3'ends...........................Poly(A)Polymerase;T4RNALigaseLigationsofDNA.................................E. coli andT4DNALigase,Ligate-ITRapidLigationKit*LigationsofRNA.................................T4RNALigaseMappingmRNA5'ends...........................S1orMungBeanNuclease;ReverseTranscriptase;RibonucleaseAMethylationofDNA..............................MethylasesMultiplexPCR...................................MagniTaqMultiplexPCRMasterMix*Nesteddeletionconstruction........................Bal 31;ExonucleaseIII;S1orMungBeanNucleaseNicktranslation..................................DNaseI;E. coli DNAPolymeraseIOligonucleotideextension..........................T7DNAPolymerase;KlenowfragmentPhosphorylation.................................T4PolynucleotideKinase,OptiKinase*Poly(A)tailingofRNA ............................Poly(A)PolymerasePolymerasechainreaction(PCR).....................TaqDNAPolymerase,RubyTaqDNAPolymerase*,FideliTaqDNAPolymerase*,HotStart-ITTaqDNA
Polymerase*Primedsynthesis.................................Klenowfragment,Exonuclease-freeKlenow*,Sequenase*QuantitativeReal-TimePCR ........................VeriQuestqPCRMasterMixes*QuantitativeReal-TimeRT-PCR......................VeriQuestOne-StepqRT-PCRMasterMixes*Replacementsynthesis............................T4andT7DNAPolymeraseRestrictionmapping..............................RestrictionendonucleasesTailingDNA ....................................TerminalTransferaseTailingRNA.....................................Poly(A)PolymeraseTranscription,promoter-specific......................E. coli, SP6,T3andT7RNAPolymerase,GeneScribeIn VitroTranscriptionKit*
Updated(*)fromTabor,S.andStruhl,K.1990“EnzymaticManipulationofDNAandRNA,”Current Protocols in Molecular Biology (F.M.Ausubel,R.Brent,R.E.Kingston,D.D.Moore,J.G.Seidman,J.A.SmithandK.Struhl,eds.),pp.3.5.15.GreenePublishingandWileyInterscience,NewYork.
Applications of Nucleic Acid Modifying Enzymes
65For bulk or alternate pack sizes, email us at [email protected].
Molecular Biology Enzymes | Binding Proteins
70028Y 200 µg70028Z 1,000 µgCustoms by request
Source:E. coli straincontaininganoverproducingcloneofE. coliRecAProtein.
Description:E. coliRecAProtein,encodedbytherecAgeneofE. coli,participatesingeneralrecombination,repairofDNAandregulationofrepair(1,2).Ithasbeenobservedin vitrotohaveDNA-dependentATPaseactivity,promoteproteolyticcleavageofrepressors(3)andtocatalyzeDNAstrandpairingandexchange(4,5).InthepresenceofATP,RecAProteinpromotesthestrandexchangeofsingle-strandDNAfragmentswithhomologousduplexDNA(6).Thisproteinwillformhelicalfilamentswithsingle-strandedandduplexDNA(7,8).
Applications:�� AnalysisofcomplexDNAstructuresbyelectronmicroscopy(9)
�� Site-directedmutagenesis(10)
�� ScreeningofDNAlibraries(11,12)
Properties:MolecularWeight:37.8kDa(13)
Purity:Greaterthan98%pureasdeterminedbySDS-PAGE.Testedforcontaminatingnon-specificendonucle-ases,exonucleases,andribonucleases.
Storage buffer:20mMTris-HCl(pH7.5),1mMDTT,0.1mMEDTA,50%glycerol.
Concentration:5mg/ml,measuredbyA280
(3)
Shipping and storage:Shippedondryice.Storeat-20°C.Forlongtermstoragestoreat-80°C.Avoidrepeatedfreeze/thawcycles.
References:1. Radding,C.M.(1978)Ann.Rev.Biochem.47,
847-880.2. Radding,C.M.(1982) Ann.Rev.Genet.16,405-
437.3. Craig,N.L.andRoberts,J.W.(1981)J. Biol.
Chem. 256,8039-8044.
4. Cox,M.M.andLehman,I.R.(1981)Proc. Natl. Acad. Sci. USA 78,3433-3437.
5. Soltis,D.A.andLehman,I.R.(1983)J.Biol.Chem.258,6073-6077.
6. Radding,C.M.(1991)J.Biol.Chem.266,5355-5358.
7. DiCapua,E.,Engel,A.,Stasiak,A.andKoller,T.(1982)J.Mol.Biol.157,87-103.
8. Register,J.C.andGriffith,J.(1986)Proc.Natl.Acad.Sci.USA83,624-628.
9. Wasserman,S.A.andCozzarelli,N.R.(1985)Proc.Natl.Acad.Sci.USA82,1079-1083.
10.Shortle,D.,Koshland,D.,Weinstock,G.M.andBotstein,D.(1980)Proc.Natl.Acad.Sci.USA77,5375-5379.
11.Honigberg,S.M.,Rao,B.J.andRadding,C.M.(1986)Proc.Natl.Acad.Sci.USA83,9586-9590.
12.Rigas,B.,Welcher,A.A.,Ward,D.C.andWeissman,S.M.(1986)Proc.Natl.Acad.Sci.USA 83,9591-9595.
13.Sancar,A.,StachelekC.,Konigsberg,W.andRupp,W.D.(1980)Proc.Natl.Acad.Sci.USA77,2611-2615.
RecA Protein
70032Y 100 µg70032Z 500 µgCustoms by request
Source:E. coli straincontaininganoverproducingcloneofE. coli SSBprotein.
Description:Single-StrandedDNABindingProteinbindswithhighaffinityinacooperativemannertosingle-strandedDNAanddoesnotbindwelltodouble-strandedDNA(1,2).ItisinvolvedinDNAreplicationandrecombinationin vivo.Thisproteinhasbeenusedtovisualizesingle-strandedDNAbyelectronmicroscopy(3,4,5).Ithasalsobeenusedinconjunc-tionwithRecAproteinforcarryingoutsite-directedmutagenesis(6)andtoselectsequencesfromlibrariesofdouble-strandedDNA(7).SSBmayalsostimulatespecificDNApolymerasesusedinDNAsequencingreactions(8)andhasbeenusedtotargetrestrictionendonucleasedigestionstospecificsitesinsingle-strandedDNAforsubsequentmutagenesis(9).
SSBhasbeenshowntobeeffectiveinfluorescencepolarizationassays(15)andeliminatespausingwhensequencingthroughstrongsecondarystructure.Morerecently,SSBwasusedtohelpobtainlongerreadlengthsinpyrosequencingforSNPanalysis(16,17).
Applications:�� EnhancementofDNApolymeraseactivity�� Fluorescencepolarizationassays(15)
�� AllowsforlongerreadlengthsinpyrosequencingforSNPanalysis(16,17)
�� Eliminatespausingwhensequencingthroughregionsofsingleanddouble-strandedDNAwithstrongsecondarystructure�� Improvesrestrictionenzymedigestions�� Site-directedmutagenesisinconjunctionwithRecA
Properties:Consistsoffouridentical18.9kDasubunits(10,11)
IsoelectricPoint:6.0(18)
NucleotidesBoundPerMonomer:8-16(18)
Thermostability:SSBretainsactivityafter20minutesincubationat65°C.
Purity:Greaterthan95%pureasdeterminedbySDS-PAGE.Testedforcontaminatingnon-specificendonucle-ases,exonucleases,andribonucleases.
Storage buffer:50mMTris-HCl(pH7.5),200mMNaCl,0.1mMEDTA,1.0mMDTT,50%glycerol.
Concentration:5µg/µl,measuredbyA280
(12)
Activity:Approximately5µgofSSBproteinisrequiredtopreventopticaldensitychangeof1µgofsingle-strandedDNAuponadditionof10mMMgCl2.
Shipping and storage:Shippedondryice.Storeat-20°C.Forlongtermstorage,storeat-80°C.Avoidrepeatedfreeze/thawcycles.
References:
1. KraussG.,Sindermann,H.,Schomburg,U.andMaass,G.(1981)Biochemistry 20,5346-5352.
2. Weiner,J.H.,Bertsch,L.L.andKornberg,A.(1975)J. Biol. Chem.250,1972-1980.
3. Chrysogelos,S.andGriffith,J.(1982)Proc. Natl. Acad. Sci. USA 79,5803-5807.
4. Sigal,N.,Delius,H.,Kornberg,T.,Gefter,M.L.andAlberts,B.(1972)Proc. Natl. Acad. Sci. USA 69,3537-3541.
5. Muskavitch,K.M.andLinn,S.(1982)J. Biol. Chem. 257,2641-2648.
6. Shortle,D.(1980)Proc. Natl. Acad. Sci. USA 77,5375-5379.
7. Honigberg,S.M.,Rao,B.J.andRadding,C.M.(1986)Proc. Natl. Acad. Sci. USA 83,9586-9590.
8. Kowalczykowski,S.C.,Bear,D.G.andVonHippel,P.H.(1981)inThe Enzymes,3rdedition,ed.P.D.Boyer,(AcademicPress,NewYork)14,373-444.
Single-Stranded DNA Binding Protein (SSB)
(Continued on next page)
66 888-362-2447 | 216-765-5000 | usb.affymetrix.com
Molecular Biology Enzymes | Binding Proteins
Standard concentration, 5 µg/µl70029Y 100 µg70029Z 500 µgHigh concentration, ≥ 10 µg/µl74029Y 300 µg74029Z 1,000 µgCustoms by request
Source:E. coli straincontaininganoverproducingcloneofT4Gene32Protein.
Description:T4Gene32Proteinisasingle-strandedDNAbind-ingproteinwhichisrequiredforT4DNAreplication,recombination,andrepair.Itbindscooperativelytosingle-strandedDNAandin vivo isrequiredinstoichiometricratherthancatalyticquantities(1,2).Italsobindstosingle-strandedRNA(with101-104loweraffinitythansingle-strandedDNA)allowingittocontrolitsownrateofsynthesisattheleveloftranslation(3,4).Ithasbeennotedtoeliminatepausingwhensequencingthroughstrongsecondarystructure.
T4Gene32ProteinhasbeenwidelyusedinstudiesofDNA-proteininteractionsandformarkingregionsofsingle-strandedDNAincytologicalpreparationsviewedbyelectronmicroscopy(5,6).Onaprimedsingle-strandedDNAtemplatetheadditionofT4Gene32Proteinresultsina5to10-foldincreaseintherateofsynthesisbyT4DNAPolymerase(7).T4Gene32ProteinhasbeenshowntobeeffectiveinstimulatingPCR.Inaddition,theenzymehasbeenfoundtoenhanceyieldandprocessivityinRNAamplification(8).
Applications:�� EnhancementofyieldandspecificityinRNAamplification(8)
�� StimulationofPCR(9)
�� Improvingrestrictionenzymedigestions(10)
�� Sequencingthroughregionsofdouble-andsingle-strandedDNApossessingstrongsecondarystructure�� EnhancementofDNApolymeraseactivity(11)
�� IncreaseyieldinRT-PCR(12,13)
Properties:IsoelectricPoint:5.5(14)MolecularWeight:33.5kDaNativeForm:MonomerNucleotidesBoundPerMonomer:10(14)
Purity:Greaterthan95%pureasdeterminedbySDS-PAGE.Testedforcontaminatingnon-specificendonucle-ases,exonucleases,andribonucleases.
Storage buffer:20mMTris-HCI(pH8.0),100mMNaCI,1.0mMEDTA,0.5mMDTT,50%glycerol.
Concentrations:PN70029:5µg/µl,measuredbyA280
(15)
PN74029:≥ 10µg/µl,measuredbyA280(15)
Functional Assay: FunctionallytestedinbindingassayofT4Gene32Proteintosingle-strandedDNAasdeterminedbyagarosegelelectrophoresisingel-shiftassay.
Shipping and storage:Shippedondryice.Storeat-20°C.
References:1. Chase,J.W.andWilliams,K.R.(1986)Ann. Rev.
Biochem.55,103-136.2. Sinha,N.K.andSnustad,D.P.(1971)J. Mol. Biol.
62,267-271.3. Krisch,H.M.,Bolle,A.andEpstein,R.H.(1974)
J. Mol. Biol.88,89-104.4. Gold,L.,O’Farrel,P.Z.andRussel,M.(1976)
J. Biol. Chem.251,7251-7262.5. Delius,H.,Mantell,N.J.andAlberts,B.(1972)
J. Mol. Biol.67,341-350.6. Brack,C.,Bickle,T.A.andYuan,R.(1975)J. Mol.
Biol.96,693-702.7. Huberman,J.A.,Kornberg,A.andAlberts,B.M.
(1971)J. Mol. Biol.62,39-52.8. Baugh,L.R.,Hill,A.A.,Brown,E.L.andHunter,
C.P.(2001)Nucl. Acids Res.29 (5),E29.9. Schwarz,K.,Hansen-Hagge,T.andBartram,C.
(1990)Nucleic. Acids Res.18,1079.10.Dombroski,D.F.andMorganA.R.(1985)J. Biol.
Chem.260,415-417.11.Topal,M.D.andSinha,N.K.(1983)J. Biol.
Chem. 258,12274-12279.12.Villalva,C.,Touriol,C.,Seurat,P.,Trempat,P.,
Delsol,G.andBrousset,P.(2001)BioTechniques31,81-86.
13.Jeffries,D.andFarquharson,C.(2002)Anal. Biochem.308,192-194.
14.Kornberg,A.andBaker,T.(1991) DNA Replication, 2ndEdition.
15.Williams,K.R.,Sillerud,L.O.,Schafer,D.E.andKonigsberg,W.H.(1979)J. Biol. Chem.254,6426-6432.
16.Shamoo,Y.,Adari,H.,Konigsberg,W.H.,Williams,K.R.andChase,J.W.(1986)Proc. Natl. Acad. Sci. USA 83,8844-8848.
T4 Gene 32 Protein
9. Milavetz,B.(1989)Nucl. Acids Res.17,3322.10.Sancar,A.,Williams,K.R.,Chase,J.W.andRupp,
W.D.(1981)Proc. Natl. Acad. Sci. USA78,4274-4278.
11.Chase,J.W.andWilliams,K.R.(1986)Ann. Rev. Biochem. 55,103-136.
12.Williams,K.R.,Spicer,E.K.,Lopresti,M.B.,Guggenheimer,R.A.andChase,J.W.(1983)J. Biol. Chem. 258,3346-3355.
13.Griffith,J.D.,Harris,L.D.andRegister,J.(1984)Cold Spring Harbor Symposia on Quant. Biol. 49,553-559.
14.Julin,D.A.,Riddles,P.W.andLehman,I.R.(1986)J. Biol. Chem. 261,1025-1030.
15.Hsu,T.M.,Chen,X., Duan,S.,Miller,R.D.and Kwok,P.-Y.(2001)BioTechniques 31,560-570.
16.Andréasson,A.,Asp,A.,Alderborn,A.,Gyllensten,U.andAllen,M.(2002)BioTechniques32 (1),124-133.
17.Ronaghi,M.(2000)Anal. Biochem286,282-288.
18.Kornberg,A.andBaker,T.(1991)DNA Replication,2ndEdition,328.
Single-Stranded DNA Binding Protein (SSB) (continued from previous page)
67For bulk or alternate pack sizes, email us at [email protected].
Molecular Biology Enzymes | Glycosylases
78310 100 unitsCustoms by request
Source: RecombinantfromGadus morhua(codliver)
Description: Uracil-DNAGlycosylase(UDGorUNG)derivedfromGadus morhua hasalltheattributesoftheenzymederivedfromE. coliwiththeaddedbenefitofbeingheat-labile.Itiscompletelyandirreversiblyinacti-vatedaftera10minuteincubationat50°C.
UDGexcisesuracilfromdU-containingDNAbycleavingtheN-glycosidicbondbetweentheuracilbaseandthesugarbackbone.Thiscleavagegener-atessitesthatareblockedfromreplicationbyDNApolymeraseorpreventedfrombecomingahybridiza-tionsite.
Double-andsingle-strandeddU-containingDNAaresubstratesforUDGwhereasRNAandnormaldT-containingDNAarenot(1).
Uracil-DNAGlycosylase(UDGorUNG)canbeutilizedforthepreventionoffalsepositiveresultsinPCRcausedbytraceamountsofcarry-overcontaminantsfromthepreviousreactions(5).ThiscanbeaccomplishedbysubstitutingdUTPfordTTPinthePCRreactionmixtoincorporatedUTPinallofthePCRproducts.
Pre-incubationofthesubsequentPCRstartingreac-tionswithUDGresultsinexcisionofuracilfromthedU-containingcarry-overcontaminantstopreventamplification.ThetemplatewhichdoesnotcontaindUisunaffectedandwillbeamplifiednormally.UDGisinactivatedduringsubsequentthermocyclingandwillnotaffectthenewdU-richPCRproducts.
Applications:�� StudyofDNArepairandmutationdetection(4)
�� IncreasecloningefficiencyofPCRproducts(2)
�� Increasetheefficiencyofsite-directedmutagenesis(3)
�� Studyofprotein-DNAinteractions(5)
�� Preventionofcarry-overcontaminantsinPCR(6)
Purity:TheenzymeischromatographicallypurifiedandhomogeneousbySDS-PAGE.Testedforcontaminat-ingendonucleasesandexonucleases.
Storage buffer:50mMTris-HCl(pH7.5),100mMNaCl,0.5mMEDTA,1mMDTT,0.1%TritonX-100,50%glycerol.
Assay conditions: Thereactionmixturecontains70mMTris-HCl,pH8.0,10mMNaCl,1mMEDTA,100µg/mlBSA,3H-dUTPlabeledDNA,andUracil-DNAGlycosylase.Incubationisat37°Cfor1hour.
Unit definition:Oneunitistheamountofenzymerequiredtoliber-ate1nmoluracilfromdU-containingDNAinonehourat37°C.
Concentration:1unit/µl
Shipping and storage:Shippedondryice.Storeat-20°C.
References:1.Lindahl,T.,Ljungquist,S.,Siegert,W.,Nyberg,B.
andSperens,B.(1977)J. Biol. Chem.252,3286-3294.
2.Nisson,P.E.,Rashtchian,A.andWatkins,P.C.(1991)PCR Methods Appl.1,120-123.
3.Kunkel,T.(1985)Proc. Natl. Acad. Sci. USA82,488-492.
4.Vaughan,P.andMcCarthy,T.V.(1998)Nucl. Acids Res.26,810-815.
5.Deuchand,P.R.,McGhee,J.D.andVanDeSande,J.H.(1993)Nucl. Acids Res.21,3437-3443.
6.Longo,M.C.,Berninger,M.S.andHartley,J.L.(1990)Gene93,125-128.
Uracil-DNA Glycosylase, Heat-Labile
71960 400 units 2,000 unitsCustoms by request
Source: E. coli straincontaininganoverproducingcloneofE. coli Uracil-DNAGlycosylase.
Description: Uracil-DNAGlycosylase(UDGorUNG)excisesuracilfromdU-containingDNAbycleavingtheN-glycosidicbondbetweentheuracilbaseandthesugarbackbone.ThiscleavagegeneratesalkalisensitiveapyrimidinicsitesthatareblockedfromreplicationbyDNApolymeraseorpreventedfrombecomingahybridizationsite.
Double-andsingle-strandeddU-containingDNAaresubstratesforUracil-DNAGlycosylasewhereasRNAandnormaldT-containingDNAarenot(1).
Applications:�� StudyofDNArepairandmutationdetection(4)
�� IncreasecloningefficiencyofPCRproducts(2)
�� Increasetheefficiencyofsite-directedmutagenesis(3)
�� Studyofprotein-DNAinteractions(5)
�� Preventionofcarry-overcontaminantsinPCR(6)
Properties:MolecularWeight:31kDaOptimumpH:8.0OptimumTemperature:37°CInactivation:95°Cfor10minutes.Note:Enzymeactivityispartiallyrestoredattem-peratureslowerthan55°C.
Purity:Testedforcontaminatingsingle-anddouble-strandedexonucleasesandendonucleases.
Storage buffer:10mMTris-HCl,pH7.5,50mMKCl,1mMDTT,0.1mMEDTA,0.05%Tween20,and50%glycerol.
Assay conditions:70mMHEPES(pH8.0),50mMEDTA,1mMDTT,0.4µglambdaDNAat37°Cfor30minutes.
Unit definition:Oneunitistheamountofenzymethatcatalyzesthereleaseof60pmolofuracilperminutefromdouble-stranded,uracil-containingDNA.
Concentration:2units/µl
Uracil-DNA Glycosylase Reaction Buffer (1 ml included):200mMTris-HCl(pH8),10mMDTT,10mMEDTA.
Shipping and storage:Shippedondryice.Storeat-20°C.
References:1.Lindahl,T.,Ljungquist,S.,Siegert,W.,Nyberg,B.
andSperens,B.(1977)J. Biol. Chem.252,3286-3294.
2.Nisson,P.E.,Rashtchian,A.andWatkins,P.C.(1991)PCR Methods Appl.1,120-123.
3.Kunkel,T.(1985)Proc. Natl. Acad. Sci. USA82,488-492.
4.Vaughan,P.andMcCarthy,T.V.(1998)Nucl. Acids Res.26,810-815.
5.Deuchand,P.R.,McGhee,J.D.andVanDeSande,J.H.(1993)Nucl. Acids Res.21,3437-3443.
6.Longo,M.C.,Berninger,M.S.andHartley,J.L.(1990)Gene93,125-128.
Uracil-DNA Glycosylase, E. coli
68 888-362-2447 | 216-765-5000 | usb.affymetrix.com
78334X 500 units78334Y 1,000 units78334Z 2,500 units
Source:E. colistraincontainingamodifiedcloneofT4PolynucleotideKinasethathasfullkinaseactivitybutlacksphosphataseactivity.
Description:OptiKinaseisamodifiedversionofT4PolynucleotideKinase(T4PNK)(1,2,3)whichprovidesgreater32Por33PlabelingefficiencyofDNA.T4PNKisahomotetra-meric,bifunctionalenzymecontaininganN-terminalkinasedomainwhichcatalyzesthetransferoftheg-phosphateofATPtothe5'hydroxylendofpoly-nucleotidesandaC-terminalphosphatasedomainwhichremovesthe3'phosphate.Inmolecularbiologyapplications,T4PNKistraditionallyusedforradiolabellingthe5'-endsofDNAandRNA.How-ever,whenphosphorylatingDNA,T4PNKdisplaysaseverebasebias,asthelabelingefficiencyisvariabledependingonthenucleotideatthe5'-end.Deoxy-guanosineatthe5'-endiskinasedwiththehighestefficiencyfollowedbydA,dT,andthendC,whichiskinasedwiththeleastefficiency(4).Unlikewild-typeT4PNK,OptiKinaseexhibitsnosuchbasebiasordis-crimination(seeFigs.1and2).Also,sinceOptiKinaselacks3'phosphataseactivity,itisparticularlyusefulforlabeling3'phosphorylatedmononucleotidesorRNAwitha3'phosphate.UseOptiKinaseforhigherlevelsofphosphorylationandforgreaterconsistencycomparedtostandardT4PNK.
Applications: �� 5'phosphorylationofsingle-strandedoligonucleotides,bothDNAandRNA�� 5'phosphorylationofdouble-strandedDNA�� EfficientlabelingofDNAwithoutabiastowardthebaseatthe5'end�� 5'phosphorylationofoligonucleotideswitha3'phosphategroup�� 5'phosphorylationofmononucleotideswitha3'phosphategroupthatcangenerateasubstance(pNp)thatcanbeligatedtothe3'endofDNAorRNAbyT4RNALigase
Properties: MolecularWeight:Homotetramerof140kDa
(4x35kDa)OptimumpH:7.6(Tris-HCI)OptimumTemperature:37°C
Purity: Greaterthan95%pureasdeterminedbySDS-PAGE.Testedforcontaminatingexonucleases,endonucle-ases,andribonucleases.
Storage buffer: 20mMTris-HCl(pH7.5),50mMNaCl,1.0mMDTT,0.1mMEDTA,50%glycerol.
Assay conditions: Thereactionmixture(100µl)contains50mMTris-HCI(pH7.6),100µMradiolabeledATP(~1.5x105cpm/nmol),10mMMgCl2,10mM2-mercaptoetha-nol,20µMspermidine,andsubstrate.Incubationisat37°C.
Unit definition: Oneunitistheamountofenzymerequiredtoincor-porate1nmolofphosphatefromradiolabeledATPintoDNAsubstratein30minutesat37°C.
Concentration: 10units/µl
Functional test: >60%phosphorylationofoligonucleotidesubstrate.
Functionally tested 10X OptiKinase Reaction Buffer (1 ml included):0.5MTris-HCI(pH7.5),100mMMgCl2,50mMDTT.
Shipping and storage: Shippedondryice.Storeat-20°C.
References: 1.Wang,L.K.andShumanS.(2001)J. Biol. Chem.
276,26868-26874.2.Wang,L.K.andShumanS.(2002)Nucl. Acids Res.
30,1073-1080.3.Richardson,C.C.(1981)TheEnzymes,3rdedition,
ed.P.D.Boyer,(AcademicPress,NewYork)14,299-314.
4.VanHouten,V.,Denkers,F.,VanDijk,M.,VanDenBredel,M.andBrakenhoff,R.(1998)Anal. Biochemistry265,386-389.
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T4 DNA LigaseStandard concentration, 1 unit/µl70005Y 100 units70005X 500 units70005 2,000 unitsHigh concentration, 10 units/µl70042X 500 units70042 2,000 units
TAE Buffer, 10X Solution75904 1 L 5 L
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TEMED76320 100 ml
Molecular Biology Enzymes | Kinases
OptiKinase(PhosphataseminusmutantofT4PNK)
Fig. 2. 5'-Phosphorylation of 5'-C oligo with varying amount of radiolabel. Test5'-Coligonucleotide18-mer(5pmol)andvary-ingamountof33P-gATPwereincubatedwith10unitsofT4PNKorOptiKinasein25μlat37°Cfor30minutes.Thespecificactivitiesoftheresulting5'-phosphorylatedoligonucleotidesweredetermined.
OptiKinase Shifts the Reaction Equilibrium Toward Phosphorylation
5'-Phosphorylation of 5'-C Oligo with Varying Amount of Radiolabel
Fig. 1. 5'-Phosphorylation of single-stranded oligonucle-otides. Testoligonucleotides*(5pmol),identicalexceptattheir5'ends,andanequimolaramountof33P-gATP,weretreatedwith10unitsofT4PNKorOptiKinasein25μlat37°Cfor30minutes.Thespecificactivitiesoftheresulting5'-phosphorylatedoligonucleotidesweredetermined.*Oliognucleotidescorrespondedto31-mers(G,A,T,andC)or18-mers(GGGandCCC).
OptiKinase Overcomes Base Bias5'-Phosphorylation of Single-Stranded Oligonucleotides
69For bulk or alternate pack sizes, email us at [email protected].
Molecular Biology Enzymes | Kinases
70031Y 500 units70031Z 1,000 units70031X 2,500 unitsCustoms by request
Source:E. colistraincontainingacloneofT4PolynucleotideKinase(1).
Description:T4PolynucleotideKinase(PNK)catalyzesthetransferoftheterminalphosphateofATPto5'hydroxylterminiofpolynucleotidessuchasDNAandRNA,oligonucleotides,and3'mononucleotides(2).InthepresenceofADP,itwillalsocatalyzetheexchangeof5'terminalphosphategroupsandATP.Italsopossessesa3'phosphatase(4,5,8,9)and2',3'cyclicphosphodiesteraseactivity(3).Thisenzymeiscom-monlyusedtolabelDNAorRNAwith32Por33Pat5'ends(6).
Applications:�� 5'-labelingofDNAandRNA�� MappingterminiofRNAtranscripts�� Phosphorylationofoligonucleotidesat5'terminipriortoligation�� Removalof3'-phosphorylgroups
Properties:MolecularWeight:140kDa(4x33kDa)OptimumpH:7.6(Tris-HCI)OptimumTemperature:37°COptimumMg2+Concentration:10mMActivators:MgCl2,spermidine,dithiothreitol,
2-mercaptoethanolInhibitors:Inorganicphosphate,pyrophosphate,
ammoniumsulfateInactivation:65°Cfor10minutes
Purity:Greaterthan95%pureasdeterminedbySDS-PAGE.Testedforcontaminatingexonucleases,endonucle-ases,andribonucleases.
Storage buffer:50mMTris-HCl(pH7.5),1.0mMDTT,0.1mMEDTA,50%glycerol.
Assay conditions:Thereactionmixture(100µl)contains50mMTris-HCI(pH7.6),100µMradiolabeledATP,10mMMgCl2,10mM2-mercaptoethanol,20µMspermi-dine,andDNAsubstrate.Incubationisat37°C.
Unit definition:Oneunitistheamountofenzymerequiredtoincorporate1nmolofradiolabeledATPintoDNAsubstratein30minutesat37°C.
Concentration:30units/µl
Functional test:Phosphorylationof10pmolofa17-meroligo-nucleotidewith20pmolradiolabeledATP,>50%incorporationoflabelwithin30minutes.
Functionally tested 10X T4 PNK Reaction Buf-fer (1 ml included):0.5MTris-HCl(pH7.6),100mMMgCl2,100mM2-mercaptoethanol.
T4 PNK Dilution Buffer (1 ml included):50mMTris-HCl(pH8.0).
Shipping and storage:Shippedondryice.Storeat-20°C.
References:1.Midgley,C.A.andMurray,N.E.(1985)EMBO J.4,
2695-2703.2.Richardson,C.C.(1981)inThe Enzymes,3rd
edition,ed.P.D.Boyer,(AcademicPress,NewYork)14,299-314.
3.Morse,D.P.andBass,B.L.(1997)Biochemistry36,8429-8434.
4.Cameron,V.andUhlenbeck,O.C.(1977)Biochemistry16,5120-5126.
5.Wang,L.K.andShuman,S.(2002)Nucl. Acids Res.30,1073-1080.
6.Maxam,A.M.andGilbert,W.(1980)Methods In Enzymology65,499-560.
7.VanHouten,V.,Denkers,F.,VanDijk,M.,VanDenBrekel,M.andBrakenhoff,R.(1998)Anal. Biochemistry265,386-389.
8.Galburt,E.,Pelletier,J.,Wilson,G.andStoddard,B.(2002)Structure10,1249-1260.
9.Wang,L.K.,Lima,C.D.andShuman,S.(2002)EMBO J.21,3873-3880.
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TEMED76320 100 ml
T4 Polynucleotide Kinase (PNK) (E.C.2.7.1.78)
70 888-362-2447 | 216-765-5000 | usb.affymetrix.com
70020Y 200 units70020Z 1,000 unitsCustoms by request
Source:E. colistraincontaininganoverproducingcloneofE. coliDNALigase.
Description:E. coliDNALigase,inthepresenceofNAD,catalyzestheformationofthephosphodiesterbondinduplexDNAcontainingcohesiveends(juxtaposed5'-phosphoryland3'-hydroxyltermini).Also,inthepresenceofcertainmacromoleculessuchasPEG,E. coliDNALigasecatalyzesbluntendligationofDNA(1).TheenzymeispurifiedbytheprocedureofPanasenko,et al.(2).E. coliDNALigasehasbeenemployedinaprocedureforhighefficiencycloningoffull-lengthcDNA(3).
Application:�� Ligationforcloning(1,3)
Properties:MolecularWeight:75kDaOptimumpH:8.1OptimumDivalentCationConcentration:
Mg2+:3mM;Mn2+:6mM
Purity:Greaterthan95%pureasdeterminedbySDS-PAGE.Testedforcontaminatingendonucleasesandexonucleases.
Storage buffer:10mMTris-HCl(pH7.5),50mMKCl,0.1mMEDTA,10mMammoniumsulfate,1mMDTT,50%glycerol.
Assay conditions:Thereactionmixture(20µl)contains30mMTris-HCl(pH8.0),4mMMgCl2,1mMDTT,26µMNAD,50µg/mlBSA,andλHindIIIfragments.Incubationisat16°Cfor30minutes.
Unit definition:Oneunitistheamountofenzymerequiredtogive50%ligationofλHindIIIfragmentsin30minutesat16°Cwitha5'-DNAterminiconcentrationof0.003µM(7µg/ml)understandardassayconditions.
Concentration:10units/µl
Functional test:LigationofλHindIIIfragments.Oneunitgives~50%ligationin30minutesat16°C.Verifiedbyagarosegelelectrophoresis.
Functionally tested 10X E. coli DNA Ligase Reaction Buffer (1 ml included):300mMTris-HCl(pH8.0),40mMMgCl2,10mMDTT,0.5mg/mlBSA,260µMNAD.
Shipping and storage:Shippedondryice.Storeat-20°C.
References:1.Zimmerman,S.B.andPheiffer,B.H.(1983)Proc.
Natl. Acad. Sci.,USA80,5852-5856.2.Panasenko,S.M.,Alazard,R.J.andLehman,I.R.
(1978)J. Biol. Chem.253,4590-4592.3.Okayama,H.andBerg,P.(1982)Mol. Cell. Biol.2,
161-170.4.Panasenko,S.M.,Cameron,J.R.,Davis,R.W.and
Lehman,I.R.(1977)Science196,188-189.
Molecular Biology Enzymes | Ligases
Standard concentration, 1 unit/µl70005Y 100 units70005X 500 units70005 2,000 unitsHigh concentration, 10 units/µl70042X 500 units70042 2,000 unitsCustoms by request
Source:E. colistraincontaininganoverproducingcloneofT4DNALigase.
Description:T4DNALigasecatalyzestheformationofaphos-phodiesterbondbetweenjuxtaposed5'phosphoryland3'-hydroxylterminiinduplexDNAorRNA.Itrepairssingle-strandednicksinduplexDNA,RNA,orDNA/RNAhybridsandwilljoinbothblunt-endandcohesive-endfragmentsofduplexDNAorRNA(1).
ForconvenienceweoffertheLigate-IT™RapidLigationKit(PN78400/10)for5minutesroomtemperatureligation.
Applications:�� Ligationofcohesive-endorblunt-endtermini�� Ligationofsyntheticlinkersoradaptors
Properties:MolecularWeight:55kDaOptimumpH:7.5-7.8OptimumTemperature:16°CforexchangereactionRequirementsforDivalentCation:Mg2+,Mn2+
OptimumMg2+Concentration:10mMSulfhydrylRequirement:2-ME,DTTStimulators:SpermineorSpermidine(0.5to1mM)Inhibitors:Na(>0.2M),K(>0.2M)Inactivation:Heatingat65°Cfor10minutes
Purity:Greaterthan95%pureasdeterminedbySDS-PAGE.Testedforcontaminatingendonucleases,exonucle-ases,andribonucleases.
Storage buffer:25mMTris-HCl(pH7.6),100mMNaCl,1mMDTT,0.5mMEDTA,50%glycerol.
Assay conditions:T4DNALigaseisassayedaccordingtothepyro-phosphateexchangemethodofWeiss,et al.Thereactionmixture(300µl)contains67mMTris-HCl,(pH7.8),6.7mMMgCl2,10mMDTT,66µMATP,and3.3µMradiolabeledpyrophosphate.Incubationisat37°Cfor20minutes.
Note:ATPisanessentialcofactorforthereaction.ThiscontrastswithE. coliDNALigasewhichrequiresNAD.
Unit definition:OneWeissunitistheamountofenzymerequiredtoconvert1nmolofradiolabeledphosphatefrompyro-phosphateintoabsorbablematerialin20minutesat37°Cunderstandardassayconditions.OneWeissunitequalsabout67cohesiveendligationunits.
Concentration:PN70005:1unit/µl;PN70042:10units/µl
Functional test:ReligationoflinearizedplasmidDNA>80%ofcontrol,asdeterminedbycountingtransformedbacterialcolonies.
Functionally tested 10X T4 DNA Ligase Reac-tion Buffer (1 ml included):660mMTris-HCl(pH7.6),66mMMgCl2,100mMDTT,660µMATP.
T4 DNA Ligase Dilution Buffer (1 ml in-cluded):20mMTris-HCl(pH7.6),60mMKCl,5mMDTT,1mMEDTA,50%glycerol.
Shipping and storage:Shippedondryice.Storeat-20°C.
References:1.Rossi,R.,Montecucco,A.,Ciarrochi,G.and
Biamonti,G.(1997)Nucl. Acids Res.25(11),2106-2113.
2.Weiss,B.,Jacquemin-Sablon,A.,Live,T.R.,Fareed,G.C.andRichardson,C.C.(1968)J. Biol. Chem.243,4543-4555.
T4 DNA Ligase (ATP-Dependent,E.C.6.5.1.1.)
E. coli DNA Ligase (NAD-Dependent,E.C.6.5.1.2.)
71For bulk or alternate pack sizes, email us at [email protected].
Molecular Biology Enzymes | Ligases
21245 600 units 2,500 unitsCustoms by request
Source:E. colistraincontaininganoverproducingcloneofT4RNALigase.
Description:T4RNALigasecatalyzesligationofa5'-phosphoryl-terminatednucleicacid(donor)anda3'-hydroxylterminatednucleicacid(acceptor)throughtheformationofa3'→5'phosphodiesterbond,withthehydrolysisofATPtoAMPandPPi(1).ItssmallestsubstratesarepNpandNpNpNanditsreactionrate(ligationefficiency)dependsonthekindsofbasesinvolved(1).InadditiontoligatingRNAtoRNA(2),itcanalsoligateDNAtoRNA,althoughslowly(3).TherateofDNAtoDNAligationisextremelyslowandthisligasealmostfailstorecognizedouble-strandednucleicacids.Anoligonucleotidewithbotha5'-phosphateand3'-OHcanserveasbothanacceptorandadonor,yieldingeithercircularormultimericproducts.
Applications:�� LigationofRNAtoRNA�� Circularizationofoligonucleotides(3)
�� Labelingthe3'terminiofRNA(4)
�� Ligationofsingle-strandedDNAtoRNA(5)
�� SpecificmodificationsoftRNAs(6)
�� CircularizationofRNA(7)
�� Cloningoffull-lengthDNA(8)
Purity:Greaterthan90%pureasdeterminedbySDS-PAGE.Testedforcontaminatingdeoxyribonucleasesandribonucleases.
Storage buffer:20mMTris-HCl(pH7.4),50mMNaCl,1.0mMDTT,1.0mMEDTA,50%glycerol.
Unit definition:Oneunitistheamountofenzymethatconverts1nmolofradiolabeledpCpintoitsacid-insolubleformin30minutesat37°C,witholigo(A)nasthesubstrate.
Concentration:10units/µl
Functional test:Ligationoftwooligonucleotides.
Functionally tested 10X T4 RNA Ligase Reac-tion Buffer (1 ml included):500mMTris-HCl(pH7.5),100mMMgCl2,100mMDTT,10mMATP,0.6mg/mlacetylatedBSA.
Shipping and storage:Shippedondryice.Storeat-20°C.
References:1.Gumport,R.I.andUhlenbeck,O.C.(1981)in
Gene Amplification and Analysis,eds.Chirikjian,J.G.andPapas,T.S.,(Elsevier,Amsterdam)2,313-335.
2.Romaniuk,P.J.andUhlenbeck,O.C.(1983)Methods in Enzymology100,52-59.
3.Brennan,C.A.,Manthey,A.E.andGumport,R.I.(1983)Methods in Enzymology100,38-52.
4.Uhlenbeck,O.C.andGumport,R.I.(1982)inThe Enzymes,3rdedition,ed.P.D.Boyer,(AcademicPress,NewYork)15,31-60.
5.Tessier,T.C.,Brousseau,R.andVernet,T.(1986)Anal. Biochem.158,171-178.
6.Bruce,A.G.andUhlenbeck,O.C.(1982)Biochemistry21,855-861.
7.White,T.C.andBorst,P.(1987)Nucl. Acids Res.15,3275-3290.
8.Dumas,J.P.,Edwards,J.B.,Delort,J.andMallet,J.(1991)Nucl. Acids Res.19,5227-5233.
T4 RNA Ligase
70440 200 units 1,000 units
Source: Recombinant
Description:T4RNALigase2,alsoknownasT4Rnl-2(gp24.1),hasbothintermolecularandintramolecularRNAstrandjoiningactivity.T4RNALigase2ismuchmoreactivethanT4RNALigase1(PN21245)injoiningnicksondouble-strandedRNAthanonjoiningtheendsofsingle-strandedRNA.Theenzymerequiresanadjacent5’phosphateand3’OHforligation.T4RNALigase2canalsoligatethe3’OHofRNAtothe5’phosphateofDNAinadouble-strandedstructure.
Applications:�� IdealforligatingnicksindsRNA�� Ligatingthe3’OHofRNAtothe5’phosphateofDNAinadouble-strandedstructure�� Suitableforligating3’OHorRNAto5’phosphateofDNAinaDNA/RNAhybrid
Purity: Greaterthan90%pureasdeterminedbySDS-PAGE.Testedforcontaminatingendonucleases,exonucle-ases,andribonucleases.
Storage buffer: 10mMTris-HCl,pH7.5,50mMKCl,35mM(NH4)2SO4,0.1mMDTT,0.1mMEDTA,50%glycerol
Assay conditions: Thereactionmixturecontains0.4μgofanequi-molarmixofa23-merand17-merRNAs,T4RNALigase2in1XT4RNALigase2ReactionBufferat37°Cin30minutes(20μlreactionvolume).
Unit definition: Oneunitisdefinedastheamountofenzymerequiredtoligate0.4μgofanequimolarmixofa23-merand17-merRNAat37°Cin30minutes.
Concentration:10units/μl
Functionally tested 10X T4 RNA Ligase 2 Reaction Buffer (1 ml included): 500mMTris-HCl,pH7.5,20mMMgCl2,10mMDTT,and4mMATP.
Storage:Shippedondryice.Storeat-20°C.
T4 RNA Ligase 2
72 888-362-2447 | 216-765-5000 | usb.affymetrix.com
14365 5 mgCustoms by request
Source:Bovinepancreas
Description:DNaseIisanendonucleasethatcatalyzesthedeg-radationofbothsingleanddouble-strandedDNAtoyielddi-,tri-,andoligonucleotideswith5'-phosphateand3'-hydroxyltermini(1-4).DNaseIisstabilizedbytheadditionofCa2+andrequiresadivalentmetalcationforactivity.Itrandomlyproducesnicksindouble-strandedDNAinthepresenceofMg2+,butinthepresenceofMn2+,bothstrandsofdouble-strandedDNAarecleaved,leavingblunt-endfragmentsorthosewithoneortwoprotrudingnucleotides(1,2).
Applications:�� LabelingofDNAbynicktranslationwithDNAPolymeraseI(PN70010Y)�� RemovalofDNAfromproteinpreps
Properties:MolecularWeight:31kDaOptimumpH:7.0,moststableatpH5.0-6.0.Activators: Mg2+andCa2+formaximumactivation. Mg2+forindependentstrandcleavage,random
sites. Mn2+forsimultaneouscleavageofbothstrandsto
yieldblunt-endedand1-2nucleotideprotrudingfragments.
Inhibitor:EDTAorEGTAHeatInactivation:75°Cfor10minutes.AddEDTA
toexcessrelativetoMg2+beforeheatingtoavoidchemicalscissionofRNA.
Purity:Testedforcontaminatingribonucleases.
Form:Lyophilizedpowderwithglycinestabilizer.
Activity:≥2,000units/mgdrybasis
Unit definition:Oneunitcausesanincreaseinabsorbanceat260nmof0.001perminutepermlat25°C,pH5.0,whenactingonhighlypolymerizedDNAinthepres-enceofionizedmagnesiumandcalcium(3).
Shipping and storage:Shippedondryice.Storeat4°C.
References:1.Bernardi,A.,Gaillard,C.andBernardi,G.(1975)
Eur. J. Biochem.52,451-457.2.Clark,P.andEichhorn,G.L.(1974)Biochemistry
13,5098-5102.3.Kunitz,M.(1950)J. Gen. Physiol.33,349-363.4.Melgar,E.andGoldthwait,D.A.(1968)J. Biol.
Chem.243,4409-4416.
Molecular Biology Enzymes | Nucleases—DNases
DNase I, Lyophilized (E.C.3.1.21.1)
14367 100 units 500 unitsCustoms by request
Source:Bovinepancreas
Description:DNaseIisanendonucleasethatcatalyzesthedegra-dationofbothsingle-anddouble-strandedDNAtoyielddi-,tri-,andoligonucleotideswith5'-phosphateand3'-hydroxyltermini(1-4).DNaseIisstabilizedbytheadditionofCa2+andrequiresadivalentmetalcationforactivity.Itrandomlyproducesnicksindouble-strandedDNAinthepresenceofMg2+,butinthepresenceofMn2+,bothstrandsofdouble-strandedDNAarecleaved,leavingblunt-endfragmentsorthosewithoneortwoprotrudingnucleotides(1,2).
Applications:�� DegradationoftemplateDNAfromRNAafterin vitrotranscription�� DeterminefootprintsofDNAbindingproteins�� NicktranslationwithDNAPolymeraseI(PN70010Y)�� RemovalofDNAfromRNApreparationspriortoRT-PCR(5)
�� GenerationofrandomoverlappingDNAlibraries(6)
Properties:MolecularWeight:31kDaOptimumpH:7.0,moststableatpH5.0-6.0.Activators: Mg2+andCa2+formaximumactivation. Mg2+forindependentstrandcleavage,random
sites. Mn2+forsimultaneouscleavageofbothstrandsto
yieldblunt-endedand1-2nucleotideprotrudingfragments.
Inhibitor:EDTAorEGTAHeatInactivation:75°Cfor10minutes.AddEDTA
toexcessrelativetoMg2+beforeheatingtoavoidchemicalscissionofRNA.
Purity:Testedforcontaminatingribonucleasesandproteases.
Storage buffer:1mMCaCl2and50%glycerol.
Unit definition:Oneunitincreasestheabsorbanceat260nmby0.001perminutepermlat25°CandpH5.0whenactingonhighlypolymerizedDNAinthepresenceofionizedmagnesiumandcalcium(3).
Concentration:Approximately2units/µl
Shipping and storage:Shippedondryice.Recommendedstoragetem-peratureis4°C;howeverthismaterialisalsostablewhenstoredat-20°C.
References:1.Bernardi,A.,Gaillard,C.andBernardi,G.(1975)
Eur. J. Biochem.52,451-457.2.Clark,P.andEichhorn,G.L.(1974)Biochemistry
13,5098-5102.3.Kunitz,M.(1950)J. Gen. Physiol.33,349-363.4.Melgar,E.andGoldthwait,D.A.(1968)J. Biol.
Chem.243,4409-4416.5.Kienzle,N.,Young,D.,Zehntner,S.,Bushell,G.and
Sculley,T.B.(1996)BioTechniques20,612-616.6.Anderson,S.(1981)Nucl. Acids Res.9,3015-
3027.
DNase I, Solution (E.C.3.1.21.1)
73For bulk or alternate pack sizes, email us at [email protected].
Molecular Biology Enzymes | Nucleases—DNases
78411 1,000 units 2,500 units
Source:Recombinant
Description:RecombinantDNaseIisoverexpressedandpurifiedfromanon-animalhost,Pichia pastoris,whichisvastlylowerinendogenousRNaselevelsthanpan-creatictissue,thesourceforbovineDNaseI.Hence,therecombinantenzymeisreadilypurifiedwithoutanydetectableRNasecontamination(Fig.1).
DNaseIisanendonucleasethathydrolyzesphosphodiesterlinkagesinDNAtoyielddi-,tri-,andoligonucleotideswitha5'-phosphateanda3'-hydroxyltermini(1).DNaseIcancleavedsDNA,ssDNA,chromatin,andDNA:RNAhybrids.However,thecleavageratesforssDNAandDNA:RNAhybridsaremuchlowerthandsDNA.ThisenzymeistotallyinactiveagainstRNA(2).Formaximumactivity,DNaseIrequiresbothCa2+andMg2+(3).Ca2+stabilizesDNaseItomaintaintheactiveconformationandMg2+isrequiredforactivity.
Applications:�� RemovalofDNAfromproteinandRNApreparations�� DegradationoftemplateDNAfromRNAafterin vitrotranscription�� RemovalofgenomicDNApriortoRT-PCR�� NicktranslationwithDNAPolymeraseI(PN70010Y)
Properties:MolecularWeight:~30kDaActivators:Mg2+andCa2+formaximumactivityInhibitors:EDTAandEGTAHeatInactivation:75°Cfor10minutesata
0.1unit/µlDNaseIconcentration.AddEDTAtoafinalconcentrationof5mMbeforeheatingtoavoidchemicalscissionofRNA.
Purity:Testedforcontaminatingribonucleasesandprote-ases.ResidualDNAcontaminationwastestedbyrealtimequantitativePCR.
Storage buffer:20mMTris-HCl,pH7.6,50mMNaCl,2mMCaCl2,2mMMgCl2,1mMDTT,0.1mg/mlpefablocSC,and50%glycerol.
Assay conditions:100µgcalfthymusDNAisincubatedin1Xreactionbufferwith40to70unitsofrDNaseIat25°C.Theabsorbanceincreaseismeasuredat260nm.
Unit definition:Oneunitistheamountofenzymethatcausesanabsorbanceincreaseof0.001perminutein1mlat260nmat25°C.
Concentration:10units/µl
10X rDNase I Reaction Buffer (1 ml included): 400mMTris-HCl,pH7.9,100mMNaCl,60mMMgCl2,10mMCaCl2.
Stop Solution (1 ml included): 50mMEDTA,pH8.0.
Shipping and storage:Shippedondryice.Storeat-20°C.
References:1.Kunitz,M.(1950)J. Gen. Physiol.33,349.2.Sutton,D.H.,Conn,G.L.,Brown,T.,and
Lane,A.N.(1997)Biochem. J.321,481-486.3.Clark,R.andEichhorn,G.L.(1974)Biochem.13,
5098.
rDNase I, RNase-Free
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TAE Buffer, 10X Solution75904 1 L 5 L
TBE Buffer, 5X Solution75891 1 L 5 L
Fig. 2. Effectiveness of rDNase I to remove DNA.1µgofhumanHeLacelltotalRNAand1µgoflambdaDNAwereco-incubatedwithout(-)orwith(+)2unitsrDNaseIin1XrDNaseIReactionBufferina10µlreactionvolume.Following15minutesat22°C,1µlofStopSolutionwasaddedandthereactionswereheat-inactivatedat65°Cfor10minutesbeforeloadingona1%agarosegel.ThisdemonstratesefficientremovalofDNAbut not RNA.
Fig. 1. Absence of RNase activity in rDNase I.2µgofhumanHeLacelltotalRNAwasincubatedwithout(-)orwith(+)2unitsrDNaseIin1XrDNaseIReactionBufferina10µlreactionvolume.Following15minutesat22°C,1µlofStopSolutionwasaddedandthereactionswereheat-inactivatedat65°Cfor10minutesbeforeloadingona1%agarosegel.TheintegrityoftheribosomalRNAindicatesthatnoRNAdigestiontookplace.
74 888-362-2447 | 216-765-5000 | usb.affymetrix.com
Molecular Biology Enzymes | Nucleases—DNases
78314 100 units 500 unitsCustoms by request
Source:Pichia pastorisstraincontainingoverproducingcloneofPandalus borealisDNase.
Description:ShrimpDNaseisanendonucleasethatcleavesphosphodiesterlinkagesinDNAtoyielddi-andoligonucleotideswith5'-phosphateand3'-hydroxyltermini.ThisDNasehasaremarkablyhighspecificactivitytowardsdouble-strandedDNA(dsDNA).TheactivitytowardsdsDNAis5,000-foldhigherthantowardssingle-strandedDNAandthuscanbeusedselectivelytodegradedsDNA,leavingsingle-strand-edDNAintact.TheactivityofthisenzymedependsonMg2+concentration(Fig.1)andisstimulatedbyCa2+.However,Ca2+alsostimulatestheRNaseactiv-ityofShrimpDNaseandshouldbeavoidedwhenRNAintegrityiscritical.
TheDNaseactivityfavorslowionicstrength.Activ-itydecreaseswithincreasingionicstrength.Thisrecombinantenzymecanbeheat-inactivatedbyamoderateheattreatmentwithouttheuseofEDTA(Fig.2).ShrimpDNaseistotallyinactivatedat70°Caftera25-30minutesincubation.
Applications:�� SelectivedegradationofdsDNAleavingssDNAandRNAintact�� RemovalofDNAfromRNApriortoRT-PCR�� RemovalofDNAtemplateafterin vitrotranscription�� NicktranslationwithDNAPolymeraseI(PN70010Y)�� FootprintdeterminationofDNAbindingprotein�� Usedinremovalofcarry-overcontaminantsinPCR(1)
Properties:MolecularWeight:47kDAOptimumpH:8.0Km(apparent):0.01mg/mlActivators:Mg2+(Optimum:10mM)andCa2+
(Optimum:1mMinthepresenceofMg2+)Inactivation:70°Cfor30minutesIonicStrength:Low(10-20mMTris-HCl)
Purity: Greaterthan98%pureasdeterminedbySDS-PAGE.Testedforcontaminatingribonucleaseandproteases.
Storage buffer: 20mMTris-HCl,pH7.5,2mMMgCl2,10mMNaCland50%glycerol.
Assay conditions:Thereactionmixturecontains100mMNaAC,pH5.0,5mMMgCl2,50µg/mlcalfthymusDNAandShrimpDNase.Incubationtemperatureis25°C(1mlreactionvolume).
Unit definition: Oneunitincreasestheabsorbanceat260nmby0.001O.D.perminuteat25°CandpH5.0usinghighmolecularweightDNAasasubstrateaccordingtothemethodofKunitz(2).
Concentration: 2units/µl
Shipping and storage:Shippedondryice.Storeat-20°C.
References:1.PCRCarryOverPreventionMethodiscoveredby
USpatent6,541,204andequivalents.AlicenseforusingthepatentedmethodisconveyedbypurchaseofShrimpDNasefromAffymetrix.
2.Kunitz,M.(1950)J. Gen. Physiol.33,349-363.
Shrimp DNase, Recombinant
Fig. 2. Residual activity of Shrimp DNase.60unitsShrimpDNasein200µlassaybufferwasincubatedat70°C.Aliquotsweretakenoutatindicatedintervalsandresidualactivitywasmeasured.ShrimpDNasewastotallyinactivatedat70°Cin25to30minutes.
Fig. 1. DNase activity as a function of [Mg2+] in Tris-buffer at pH 8.0.Minimumof2mMmagnesiumwasrequiredforDNaseactivity.Highestactivitywasobservedat10mM(datanotshown).
78331 1 ml
Description: RecommendedforuseintheProkaryoticTargetPreparationprotocolfromAffymetrixfortheGeneChip®P. aeruginosaGenomeArrayandtheGeneChipE. coliAntisenseGenomeArray.
10X DNase I Buffer Formulation: 100mMTrisAcetate,pH7.5,100mMMagnesiumAcetate,and500mMPotassiumAcetate
Storage:4°C
10X DNase I Buffer
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RT-PCR Master Mix (2X)78370 100 reactions
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T7 RNA PolymeraseStandard concentration, 20 units/μl70047Y 6,000 unitsHigh concentration, >80 units/μl70001Z 30,000 units
75For bulk or alternate pack sizes, email us at [email protected].
Molecular Biology Enzymes | Nucleases—Endonuclease
78300 50,000 unitsCustoms by request
Source:E. coli straincontaininganoverproducingcloneoftheT4EndonucleaseVIIprotein.
Description:T4EndonucleaseVIIisaDNAjunctionspecificendo-nuclease(alsoreferredtoasaresolvaseoracleav-ase).T4EndonucleaseVIIistheproductofgene49ofphageT4.Thisgeneproducthas157aminoacidresiduesandamassof18kDa.T4EndonucleaseVIIfunctionsin vivotoresolvebranchedDNAstructuresinnewlysynthesizedDNA.ItdoesthisbycleavingtheDNAflankingthebranchpriortorearrangementandreannealingoftheDNAstrands.T4Endonucle-aseVIIalsorecognizesmismatches,insertionordeletionloops,gaps,andapurinic/apyrimidinicsites,andcreatesnicksintheDNAstrandsatthesesites(1).ThewidevarietyofsubstratesacceptedsuggeststhatT4EndonucleaseVIIrecognizeschangesinthestructureorconformationofDNAratherthanbind-ingtoaspecificsequence(2).T4EndonucleaseVIIhasbeencrystallizedasadimer(3),althoughthenumberofsubunitsrequiredforbranchresolutionisunclear,asmultiplesofdimershavebeenreportedtobindDNAbranchstructuresduringin vitroDNAbindingexperiments(4).
Applications:�� SNPanalysis(5,6)
�� Mutationdetection(5,6)
Storage buffer:20mMTris-HCI(pH8.0),5mMDTT,50%glycerol.
Concentration:500units/µl
Functional assay: 1500units(15ng)ofT4EndonucleaseVIIdegrades50%of100fmolTAMRAlabeleddouble-stranded24-meroligonucleotidesubstratein15minutesat37°Cin50mMTris-HCl,pH8.0,10mMMgCl2,10mMβ-MEand0.5µg/µlBSA.
T4 Endonuclease VII Dilution Buffer (1 ml included): 10mMTris-HCI(pH7.5),1mMEDTA,50%glycerol.
Shipping and storage:Shippedondryice.Storeat-20°C.
References:1.Kemper,B.(1997)inDNA Damage and Repair,
eds.Nickoloff,J.A.,Hoekstra,M.(HumanaPress,Totowa,NJ),1,179-204.
2.Greger,B.andKemper,B.(1998)Nucl.Acids Res.26,4432-4438.
3.Raaijmakers,H.,Vix,O.,Toro,I.,Golz,S.,Kemper,B.andSuck,D.(1999)Embo J.,18,1447-1458.
4.Kupfer,C.,Lee,S.andKemper,B.(1998)J.Biol.Chem.273,31637-31639.
5.Youil,R.,Kemper,B.andCotton,R.G.(1995)Proc. Natl. Acad. Sci. USA 92,87-91.
6.Golz,S.andKemper,B.(1999)Nucl.Acids Res.27,e7.
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T4 Endonuclease VII (T4gp49,HJResolvase)
78454 1,000 units 5,000 units
Source:AnE. colistrainwhichcarriestheclonedhumanAPE1gene.
Description: Humanapurinic/apyrimidic(AP)endonuclease(APE1)cleavesthephosphodiesterbackboneaDNAstrandimmediately5'toanAPsiteviaahydrolyticmechanism.Cleavagegeneratesasingle-strandDNAbreakwitha3'-hydroxyland5'-deoxyribosephosphateterminus.Inadditiontoendonucleaseactivity,APE1hasbeenreportedtohaveweakDNA3'-diesterase,3'to5'exonuclease,andRNaseHactivities.APE1shareshomologywithEscherichia coliexonucleaseIIIprotein,andisalsoreferredtoasHAP1orRef-1.
Applications:�� Singlecellgelelectrophoresis(cometassay)�� Alkalineelution�� Alkalineunwinding�� Modifiednicktranslation�� Mismatchrepair
Purity: Testedforcontaminatingendonucleases,exonucle-ases,andribonucleases.
Storage buffer:10mMTris-HCl(pH7.5),50mMKCl,1mMDTT,0.1mMEDTA,0.05%Tween-20,50%Glycerol
Concentration:10units/µl
Unit definition:Oneunitisdefinedastheamountofenzymerequiredtocleave20pmolofanoligonucleotideduplexcontainingasingleAPsiteinatotalreactionvolumeof10µlin1hourat37°C.AnAPsiteiscreatedbytreating20pmolofanoligonucleotideduplexcontainingasingleuracilresiduewith1unitofE.coliUracil-DNAGlycosylase(UDGorUNG,PN71960)for2minutesat37°C.
10X APE 1 Reaction Buffer (1 ml included):200mMTris-acetate(pH8.0),500mMpotassiumacetate,100mMmagnesiumacetate,10mMDTT.
Storage:Shippedondryice.Storeat-20°C.
References:1.Demple,B.et al.(1991)Proc. Natl. Acad. Sci. USA
88,11450-11454.2.Barzilay,G.et al.(1995)Nucl. Acids Res.,23,
1544-1550.3.Barzilay,G.et al.(1995)Nature Struc. Biol.2,
451-468.4.Robson,C.N.andHickson,D.I.(1991)Nucl. Acids
Res.19,5519-5523.5.Flaherty,D.M.(2001)Am. J. Respir. Cell. Mol. Biol.
25,664-667.
Human Apurinic/Apyrimidinic Endonuclease 1 (APE 1)
76 888-362-2447 | 216-765-5000 | usb.affymetrix.com
Standard concentration, 10 units/µl70073Z 2,500 units70073X 5,000 unitsHigh concentration, 20 units/µl72073 5,000 unitsCustoms by request
Source:E. colistraincontaininganoverproducingcloneofE. coliExonucleaseI.
Description:ExonucleaseIhydrolyzessingle-strandedDNAinthe3'→5'direction,releasing5'-mononucleotidesandleavingtheterminal5'-dinucleotideintact.Hydrolysisisprocessiveandcannotproceedifthe3'terminusisphosphorylated.ExonucleaseIcanbeusedtomeasuretheendonucleolyticcleavageofcovalentlyclosedcircularsingle-strandedDNAreactedwithanendonucleaseofinterest(1).Inaddition,DNAhelicaseactivitycanbemeasuredutilizingExonucleaseI(2).
ExonucleaseIisparticularlyusefulinpreparingtheproductsofPCRforapplicationsinvolvingsequencingorlabelingmethods(3).Typically,theexcessprimersandanyotherextraneoussingle-strandedDNApresentinPCRproductswillinterferewithsubsequentenzymaticreactionsinvolvingDNAsynthesis.ThehydrolyticpropertiesofExonucleaseIdegradeallsingle-strandedDNApresentinthePCRmixtureallowingtheproducttobeusedmoreefficientlyinotherapplications.WhencombinedwithShrimpAlkalinePhosphatase(PN78390)fordNTPdephosphorylation,theuseofalternativepurificationmethods,suchascolumns,gelsormagneticseparations,arecompletelyeliminated.
Applications:�� Eliminationofresidualsingle-strandedDNAcontaininga3'terminus�� MeasuringendonucleolyticcleavageofcovalentlyclosedcircularssDNA�� MeasuringDNAhelicaseactivity
Properties:MolecularWeight:55kDaHeatInactivation:80°for15minutesDegradestoterminaldinucleotidesDegradesglycosylatedDNAOptimumTemperature:37°C
Purity:Greaterthan95%pureasdeterminedbySDS-PAGE.Testedforcontaminatingendonucleases,double-strandedexonucleases,andribonucleases.
Storage buffer:20mMTris-HCI(pH7.5),5mM2-mercaptoethanol,0.5mMEDTA,50%glycerol.
Assay conditions:Thereactionmixture(100µl)contains67mMglycinebuffer(pH9.5),10mM2-mercaptoetha-nol,6.7mMMgCl2,0.5mMdenaturedDNA,andenzyme.Incubationisat37°Cfor30minutes.
Unit definition:Oneunitistheamountofenzymewhichcatalyzesthereleaseof10nmolofacid-solublenucleotidefromdenaturedDNAin30minutesat37°Cunderstandardconditions.
Concentration:PN70073Z/X:10units/µl;PN72073:20units/µl
Functional assay:TreatedPCRproductwithExonucleaseItodegradeunincorporatedprimersbeforesequencingwithSequenaseVersion2.0DNASequencingKit(PN70770).
Shipping and storage:Shippedondryice.Storeat-20°C.
References:1.Goldmark,P.J.andLinn,S.(1972)J. Biol. Chem.
247,1849-1860.2.Rosamond,J.,Telander,K.M.andLinn,S.(1979)
J. Biol. Chem.254,8646-8652.3.Werle,E.,ScneiderC.,Renner,M.,Völker,M.and
Fiehn,W.(1994)Nucl. Acids Res.22,4354-4355.4.Hanke,M.andWink,M.(1994)BioTechniques
17,858-860.
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Exonuclease I
Molecular Biology Enzymes | Nucleases—Exonucleases
Exonuclease comparison
Exonuclease Gene(s) DNA substrates Mode of action Acid-soluble products
ExonucleaseI sbcB(xonA) Single-stranded 3'→5' 5'-dNMP5'-terminalpNpN
ExonucleaseIII xthA Duplex 3'→5' 5'-dNMP
ExonucleaseVII xseA, xseB Single-stranded3'→5'5'→3' Oligonucleotides
T7Gene6Exonuclease T7 gene 6 Duplex 5'→3' Oligonucleotides5'-dNMP
77For bulk or alternate pack sizes, email us at [email protected].
70023Y 5,000 units70023Z 25,000 unitsCustoms by request
Source:E. colistraincontaininganoverproducingcloneoftheE. coli xthAgene(ExonucleaseIII).
Description:ExonucleaseIII,themajorapurinic/apyrimidinic(AP)endonucleaseinE. coli(1),isa3'-5'exonucleasespecificfordsDNA.Thisenzymecatalyzesthestep-wiseremovalof5'-mononucleotidesfromthe3'-endsofdsDNAbutnotfromssDNA.ExonucleaseIIIisactiveondsDNAwithbluntends,5'-overhangs,andnicks,butnotonprotruding3'-ends4basesorlonger(2).Thesepropertiesareutilizedtofacilitatestrand-specificlabeling,unidirectionaldeletions,andselectivestranddegradation.ExonucleaseIIIalsohasseveralotheractivitiesincluding:digestionoftheRNAstrandofanRNA-DNAheteroduplex(3),strand-cleavageonthe5'sideofAPsitesinbothdsDNAandssDNA(4),removalof3'-phosphatesfromdsDNA(5),andanincreaseinMutYturnover(6).
Applications:�� Preparingstrand-specificradioactiveprobeswithaDNApolymerase�� Preparingsingle-strandedDNAtemplatesfordideoxysequencing(3)
�� Constructingunidirectionaldeletions�� LongPCR�� DNAfootprinting�� DNArepair/mutationdetectionstudies
Properties:MolecularWeight:32kDaInactivation:Heatat75°Cfor10minutesoradd8µl
0.5MEDTAina200µlreactionvolume.OptimumpH:7.6OptimumTemperature:37°COptimumMg2+Concentration:5mM
Purity:Greaterthan99%pureasdeterminedbySDS-PAGE.Testedforcontaminatingendonucleasesandribonuclease.
Storage buffer:50mMTris-HCl(pH8.0),1mMDTT,100mMKCl,50%glycerol.
Assay conditions:Thereactionmixture(200µl)contains50mMTris-HCl(pH8.0),10mM2-mercaptoethanol,5mMMgCl2,activatedDNA,andenzyme.Incubationisat37°Cfor30minutes.
Unit definition:Oneunitwillcatalyzethereleaseof1nmolofnucleotidestoacidsolubleformin30minutesat37°Cunderstandardassayconditions.
Concentration:200units/µl
Functional test:Productionofaseriesofdeletionsfromalinearizedplasmid,witha4base3'protrusionatoneendanda5'overhangorbluntendattheother.
Functionally tested 10X Exonuclease III Reaction Buffer (1 ml included):660mMTris-HCl(pH8.0),66mMMgCl2,50mMDTT.
Shipping and storage:Shippedondryice.Storeat-20°C.
References:1.Taylor,A.F.andWeiss,B.(1982)J. Bact.151(1),
351-357.2.Henikoff,S.(1984)Gene28,351-359.3.Linn,S.M.(1982)inLinn,S.M.andRoberts,R.J.
ed.Nucleases,291-309,ColdSpringHarborLaboratoryPress.
4.Shida,T.,Noda,M.andSekijuchi,J.(1996)Nuc. Acids Res.24(22),4572-4576.
5.Doetsch,P.W.andCunningham,R.P.(1990)Mutat Res.236(2-3),173-201.
6.Pope,M.A.,Porello,S.andDavid,S.(2002)J. Biol. Chem.277(25),22605-22615.
Exonuclease III (E.C.3.1.11.2)
70082Y 200 units70082Z 1,000 unitsCustoms by request
Source:E. colistraincontainingoverproducingclonesencod-ingboththelarge(xseA)andsmall(xseB)subunitsofExonucleaseVII.
Description:ExonucleaseVIIisastrictsingle-stranddirecteden-zymewith5'→3'and3'→5'exonucleaseactivities,makingittheonlybi-directionalexonucleasewithsingle-strandedspecificity(1,2).ExonucleaseVIIhasnoapparentrequirementfordivalentcations,asitisfullyactiveinthepresenceofEDTA.Initialreactionproductsareacidinsolubleoligonucleotideswhicharefurtherhydrolyzedintoacidsolubleform.Theproductsoflimiteddigestionsaresmalloligomers(dimerstododecamers).
Applications:�� MappingpositionsofintronsingenomicDNA(3)
�� RemovalofvectorDNAfrominsertswithpoly(dA-T)tails(4)
�� RemovalofexcessPCRprimers(5)
�� Removalofsingle-strandedover-hangsproducedbyrestrictionendonucleases
Properties:MolecularWeight: xseAsubunit=51.8kDa; xseBsubunit=8.8kDaOptimumTemperature:37°COptimumpH:8.0Inactivation:95°Cfor10minutes
Purity:Greaterthan95%pureasdeterminedbySDS-PAGE.Testedforcontaminatingendonucleases,double-strandedexonucleases,andribonucleases.
Storage buffer:50mMTris-HCl(pH8.0),200mMNaCl,0.5mMEDTA,10mMDTT,50%glycerol.
Assay conditions:Thereaction(50µl)contains50mMTris-HCl(pH7.9),50mMpotassiumphosphate(pH7.6),8.3mMEDTA,10mM2-mercaptoethanol,denaturedDNA,andenzyme.
Unit definition:Oneunitistheamountofenzymerequiredtocatalyzetheconversionof1nmolofnucleotidetoacid-solublenucleotidein30minutesat37°Cunderstandardassayconditions.
Concentration:10units/µl
Shipping and storage:Shippedondryice.Storeat-20°C.
References:1.ChaseJ.W.andRichardson,C.C.(1974)J. Biol.
Chem.249,4545-4552.2.ChaseJ.W.andRichardson,C.C.(1974)J. Biol.
Chem.249,4553-4561.3.Berk,A.J.andSharp,P.A.(1978)Proc. Natl. Acad.
Sci. USA75,1274-1278.4.Goff,S.P.andBerg,P.(1978)Proc. Natl. Acad. Sci.
USA75,1763-1767.5.Li,H.H.,Cui,X.andArnheim,N.(1991)Nucl.
Acids Res.19,3139-3141.
Exonuclease VII
Molecular Biology Enzymes | Nucleases—Exonuclease
78 888-362-2447 | 216-765-5000 | usb.affymetrix.com
Molecular Biology Enzymes | Nucleases—Exonucleases
70025Y 2,000 units70025Z 10,000 unitsCustoms by request
Source:E. coli straincontaininganoverproducingcloneofT7Gene6Exonuclease(1).
Description:T7Gene6ExonucleasehydrolyzesduplexDNAnon-processivelyinthe5'→3'directionfromboth5'-phosphorylor5'-hydroxylnucleotidesbyliberat-ingoligonucleotides(2)aswellasmononucleotidesuntilabout50%oftheDNAisacidsoluble(1,3).Italsodegradesnucleotidesatthegapsandnicksofdouble-strandedDNAfromthe5'-terminiandRNAfromRNA:DNAhybridsinthe5'→3'direction.
T7Gene6ExonucleaseissimilartoLambdaExonu-cleaseinthatitcatalyzesthestepwisehydrolysisofduplexDNAfromthe5'terminiliberating5'mono-nucleotides.However,unlikeLambdaExonuclease,theenzymehaslowprocessivityanditwillremoveboth5'-hydroxyland5'-phosphoryltermini(4).
ItalsodegradesRNAandDNAfromRNA:DNAhy-bridsinthe5'→3'directionbutisunabletodegradeeitherdouble-orsingle-strandedRNA.
Applications:�� Controlledstepwisedigestionofdouble-strandedDNAfromthe5'termini�� GeneratingssDNAtemplatesforsequencing(5)orSNPanalysis(6)
Properties:MolecularWeight:32kDaOptimumpH:7.5OptimumTemperature:37°CInactivation:75°Cfor10minutesoradd2µlof
0.5MEDTAfora50µlreactionvolume.
Purity:Greaterthan95%pureasdeterminedbySDS-PAGE.Testedforcontaminatingendonucleasesandribonucleases.
Storage buffer:50mMKPO4(pH6.5),1mMEDTA,1mMDTT,50%glycerol.
Assay conditions:Thereactionmixture(50µl)contains50mMTris-HCI(pH8.1),5mMMgCl2,20mMKCI,5mM2-mercap-toethanol,double-strandedDNA,andenzyme.Incubationisat37°Cfor15minutes.
Unit definition:Oneunitistheamountofenzymerequiredtorelease1nmolofacidsolublenucleotidein15minutesat37°Cunderstandardassayconditions.
Concentration:50units/µl
Functional test:Conversionof0.5pmolofλDNAtosingle-strandedhalfmoleculesby75unitsofenzymein30minutesat37°C.Verificationbyagarosegelelectrophoresis.
Functionally tested 5X T7 Gene 6 Exonuclease Reaction Buffer (1 ml included):200mMTris-HCl(pH7.5),100mMMgCl2,250mMNaCl.
Shipping and storage:Shippedondryice.Storeat-20°C.
Digestion protocolDigestion of double-stranded DNA with T7 Gene 6 ExonucleaseDNA(~2.5μg/μl) ___μl5XT7Gene6ExonucleaseBuffer 3μlT7Gene6Exonuclease(5units/μgDNA) ___μlWater to15μl
Incubateat37°Cfor30minutes.
Incubateat80°Cfor10minutestoinactivatetheenzyme.
Productsshouldbecheckedonanagarosegel.ToavoidtrappingofDNAinthewellsofagarosegelsor“sticky-end”artifactbands,heatsamplesto65–75°Cfor3–4minutespriortoloading.
Note: Theabovebufferisa“mediumsalt”bufferwhichworksformanyrestrictionenzymesbutmaynotworkforallofthem.Ifaspecificbufferisusedfordigestion,subsequentdigestionwithT7Gene6Exonucleaseusuallystillworkssatisfactorily.Subsequentreactionsshouldbecarriedoutintheappropriatebuffer.Ethanolprecipitationafterdiges-tionmaybeusedtochangebuffers.
References:1.Kerr,C.andSadowski,P.D.(1972)J. Biol. Chem.
247,311-318.2.Kornberg,A.andBaker,T.(1991)DNAReplication,
SecondEdition,591.3.Thomas,K.R.andOlivera,B.M.(1978)J. Biol.
Chem.253,424-429.
4.Ausubel,F.M.,Brent,R.,Kingston,R.E.,Moore,D.D.,Seidman,J.G.,Smith,J.A.andStruhl,K.,(1987)Current Protocols in Molecular Biology (JohnWileyandSons,Inc.).
5.Shon,M.,Germino,J.andBastia,D.(1982)J. Biol. Chem.257,13823-13827.
6.Nikiforov,T.T.,Rendle,R.B.,Goelet,P.,Rogers,Y.H.,Kotewicz,M.L.,Anderson,S.,Trainor,G.L.andKnapp,M.R.(1994)Nucl. Acids Res22,(20),4167-4175.
7.Engler,M.J.andRichardson,C.C.(1983)J. Biol. Chem.258,11197-11205.
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ExoSAP-IT PCR Product Cleanup78250 20 reactions78200 100 reactions78201 500 reactions78202 2,000 reactions78205 5,000 reactions
HT ExoSAP-IT High-Throughput PCR Product Cleanup78395 480 rctns x 8-tube strip 5,760 rctns x 1 plate (12 x 8-tube strips) 23,040 rctns - 4 plates 1,000 rctns (2 ml) 5,000 rctns (10 ml)
T7 Gene 6 Exonuclease
79For bulk or alternate pack sizes, email us at [email protected].
27-0330-02 1 gmCustoms by request
Source:Bovinepancreas
Description:RNaseIcatalyzesthehydrolysisof3'→5'phos-phodiesterbondsofRNAwiththeformationofoli-goribonucleotidesterminatinginpyrimidine2'→3'cyclicphosphate.RNaseIisapreparationcontainingamixtureof70%RNaseA.Theremaining30%consistsofotherisozymesofRNase.
Applications:�� RemovalofRNAfromDNA�� RemovalofRNAfromproteinpreparations(2)
Form:Lyophilizedpowder
Properties:Inhibitors:Ribonucleaseinhibitorandvanadyl-
ribonucleosidecomplexes.Inactivation:70°Cfor15minutes
Purity:Theenzymeischromatographicallypurifiedandistestedforcontaminatingproteases.
Unit definition:One(Kunitz)unitistheamountofenzymewhich
catalyzesthehydrolysisofyeastRNAtoyieldafirstordervelocityconstantof1.0at25°C,pH5.0(1).
Concentration:Approximately70Kunitzunits/mg
Shipping and storage:Shippedondryice.Storeat-20°C.
References:1.Asubel,F.M.,Brent,R.,Kingston,R.E.,Moore,
D.D.,Seidman,J.G.,Smith,J.A.andStruhl,K.(1998)Current Protocols in Molecular Biology(JohnWileyandSons,Inc.).
2.Kalintsky,G.,Hummel,J.P.andDierks,C.(1959)J.Biol.Chem.234,1512-1516.
RNase I
Molecular Biology Enzymes | Nucleases—RNases
78020Y 100 mgCustoms by request
Source:Bovinepancreas
Description:RNaseAspecificallycleavessingle-strandedRNAat3'phosphatelinkagesofpyrimidineresiduesleavingpyrimidine3'phosphatesandoligonucleotideswithterminalpyrimidine3'phosphates(1).
Applications:�� CleavingunhybridizedareasofRNAfromRNA:DNAhybridsinRNAorDNAmapping(2)
�� RemovingcontaminatingRNAfromDNAmini-preps.(Add1µlofa10mg/mlRNaseAsolutiontoresuspendedDNApelletpreparedfroma1.5mlbacterialculture)
Form:Lyophilizedpowder
Properties:MolecularWeight:13.7kDaInhibitors:Ribonucleaseinhibitor;vanadyl-
ribonucleosidecomplexes
Purity:TheenzymeischromatographicallypurifiedandistestedforcontaminatingproteasesandDNases.
Assay conditions:TwounitsofRNaseAareincubatedwithTorulayeastRNAin0.05Msodiumacetate,pH5.0,inavolumeof3mlat25°C.Absorbanceat300nmisreadat15secondintervalsfor4minutes;incubationthencon-tinuesat25°C.Absorbanceistakenat300nmagainafter3.5hoursoruntilasteadystateisobtained.
Unit definition:OneunitistheamountofenzymerequiredtocatalyzethehydrolysisofRNAataratesuchthatK(velocityconstant)equalsunity(Kunitzunits)atpH5.0and25°C.
Concentration:Approximately80units/mg
Shipping and storage:Shippedondryice.Storeat-20°C.
References:1.Davidson,J.N.(1972) The Biochemistry of the
Nucleic Acids,7thedition,(AcademicPress,NewYork).
2.Myers,R.M.,Larin,Z.andManiatis,T.(1985)Science230,1242-1246.
RNase A, Lyophilized (E.C.3.1.27.5)
27-0323-01 100 mg
Source: Bovinepancreas
Description: RNaseI‘A’specificallycleavessingle-strandedRNAat3’phosphatelinkagesofpyrimidineresiduesleav-ingpyrimidine3’phosphatesandoligonucleotideswithterminalpyrimidine3’phosphates(1).
Applications: �� CleavingunhybridizedareasofRNAfromRNA:DNAhybridsinRNAorDNAmapping(2).�� RemovingcontaminatingRNAfromDNAmini-preps.(Add1μlofa10mg/mlRNaseAsolutiontoresuspendedDNApelletpreparedfroma1.5mlbacterialculture.)
Form: Lyophilizedpowder
Properties: MolecularWeight:13.7kDa(RNaseA),14±0.3
kDa(RNaseB)pI:9.45OptimumpH:7.0-7.5Inhibitors:Heavymetalions,RNaseinhibitors,
vanadyl-ribonucleosidecomplexes.Inactivation: InactivationofRNaseIcanbespecifi-callyachievedusingaRNaseinhibitor.RemovalofRNaseinhibitoristypicallyachievedbytreatmentwithProteinaseKfollowedbyphenolextractionsandethanolprecipitations.
Purity: RNaseIcontains70%RNaseAandtheremaining30%isotherisozymesofRNase.ThisRNaseIisasaltfree,chromatographicallypreparedfreezedriedpowder. Testedforproteaseactivity.
Unit definition: OneKunitzunitcatalyzesthehydrolysisofyeast
RNAtoyieldafirst-ordervelocityconstantof1.0at25°C,pH5.0.
Concentration: ≥70Kunitzunits/mg
Notes: Activeunderawiderangeofconditions.Underlow-saltconditions(0-100mMNaCl),single-strandedRNA,double-strandedRNA,andtheRNAstrandofanRNA:DNAduplexarecleaved.Withhighsalt(>300mMNaCl),single-strandedRNAiscleaved.
Shipping and storage: Shippedondryice.Storeat-20°C.
References:1.Davidson,J.N.(1972)The Biochemistry of the
Nucleic Acids,7thedition,(AcademicPress,NewYork).
2.Myers,R.M.,Larin,Z.,andManiatis,T.(1985)Science,230,1242-1246.
RNase I ‘A’, Powder
80 888-362-2447 | 216-765-5000 | usb.affymetrix.com
Molecular Biology Enzymes | Nucleases—RNases
70194Y 1 mg70194Z 5 mgCustoms by request
Source:Bovinepancreas
Description:RNaseAspecificallycleavessingle-strandedRNAat3'phosphatelinkagesofpyrimidineresiduesleavingpyrimidine3'phosphatesandoligonucleotideswithterminalpyrimidine3'phosphates(1).Thisenzymedoesnotrequireco-factorsanddivalentcationsfortheactivity.
Applications:�� CleavingunhybridizedareasofRNAfromRNA:DNAhybridsinRNAorDNAmapping(2)
�� RemovingcontaminatingRNAfromDNAmini-preps.(Add30unitsRNaseAsolutiontoresuspendedDNApelletpreparedfroma1.5mlbacterialculture)�� Preparationofrecombinantproteins�� Ribonucleaseprotectionassays(3)
�� PlasmidandgenomicDNAisolation(4,5)
�� Mappingsingle-basemutationsinDNAorRNA(6,7)
Properties:MolecularWeight:13.7kDaInhibitors:Ribonucleaseinhibitor;vanadyl-
riboncleosidecomplexes;heavymetalionsOptimumpH:7.0-7.5
Purity:TheenzymeischromatographicallypurifiedandistestedforproteaseandDNaseactivities.Itisnotnecessarytoboiltheenzymebeforeuse.
Storage buffer:50mMsodiumacetate,(pH5.0)0.3mMEDTA,50%glycerol.
Unit definition:OneunitofRNaseAistheamountofenzymerequiredtocauseanincreaseinabsorbanceof1.0at260nmat37°C(pH5.0)whenyeastrRNAishydrolyzedtoacid-solubleoligonucleotides.OneKunitzunitisequivalentto50USBunits.
Concentration:≥ 20units/µl;~5mg/ml
Shipping and storage:Shippedondryice.Storeat4°Cor-20°C.
References:1.Davidson,J.N.(1972) The Biochemistry of the
Nucleic Acids,7thedition,(AcademicPress,NewYork).
2.Myers,R.M.,Larin,Z.andManiatis,T.(1985)Science230,1242-1246.
3.Sambrook,J.,Russell,D.W.(2001)Molecular Cloning: A Laboratory Manual,thethirdedition,ColdSpringHarborLaboratoryPress,ColdSpringHarbor,NewYork,7.63-7.74.
4.Sambrook,J.,Russell,D.W.(2001)Molecular Cloning: A Laboratory Manual,thethirdedition,ColdSpringHarborLaboratoryPress,ColdSpringHarbor,NewYork,1.31-1.38.
5.Sharma,R.C.,MurphyA.J.,DeWald,M.G.andSchimke,R.T.(1993)BioTechniques14,176-178.
6.MyersR.M.,Larin,Z.andManiatis,T.(1985)Science230,1242-1246.
7.WinterE.,Yamamoto,F.,Almoguera,C.andPerucho,M.(1985)Proc. Natl. Acad. Sci.USA,82,7575-7579.
RNase A, Molecular Biology Grade, Solution (E.C.3.1.27.5)
70054Y 250 units70054Z 1,000 unitsCustoms by request
Source:E. colistraincontaininganoverproducingcloneofE. coliRNaseH.
Description:E. coliRNaseHisanendoribonucleasethatdegradestheRNAportionofDNA:RNAhybrids(1).ItisakeyenzymeintheOkayama-BergandGubler-HoffmancDNAsynthesisandcloningprocedures,inwhichitisemployedtoremovethemRNAduringsecond-strandcDNAsynthesis(2,3).RNaseHisusefulfortheremovalofpoly(A)tailsonmRNAsafterhy-bridizationwitholigo(dT)(4-6).IthasalsobeenusedforthespecificfragmentationofRNAafterhybridiza-tionoftheRNAwitholigodeoxynucleotides(7).
Applications:�� RemovesthemRNAstrandoftheRNA:DNAduplexduringsecondstrandcDNAsynthesis�� Removalofpoly(A)tailsonmRNAafterhybridizationwitholigodT�� MakingspecificfragmentationofRNAafterhybridizationofRNAwitholigodeoxynucleotides�� Site-specificcleavageofRNA(7)
�� Studyofin vitropolyadenylationreactionproducts(8)
Properties:Activators:Mg2+orMn2+
MolecularWeight:21kDa
Purity:Testedforcontaminatingnon-specificendonucle-ases,exonucleases,andribonucleases.
Storage buffer:20mMTris-HCl(pH7.9),100mMKCl,10mMMgCl2,0.1mMEDTA,0.1mMDTT,50%glycerol.
Assay conditions:Reaction(100µl)contains20mMTrisHCl(pH8.0),100mMKCl,10mMMgCl2,1mMDTT,0.5nmolpoly(rA):poly(dT),enzyme.
Unit definition:Oneunitcatalyzesthehydrolysisof1nmolofRNAinradiolabeledpoly(A)-poly(dT)toacid-solublematerialin20minutesat37°C.
Concentration:5units/µl
5X RNase H Reaction Buffer (1 ml included):100mMTris-HCl(pH7.5),100mMKCl,50mMMgCl2,0.5mMEDTA,0.5mMDTT
Shipping and storage:Shippedondryice.Storeat-20°C.
References:1.Crouch,R.J.(1981)in Gene Amplification and
Analysis,eds.J.G.ChirikjianandT.S.Papas,(Elsevier/North Holland, Inc., New York)2,217-228.
2.Okayama,H.andBerg,P.(1982)Mol.Cell Biol.2,161-170.
3.Gubler,U.andHoffman,B.J.(1983)Gene 25,263-269.
4.Vournakis,J.N.,Efstratiadis,A.andKafatos,F.C.(1975)Proc.Natl.Acad.Sci.USA72,2959-2963.
5.Narayan,P.,Ayers,D.F.,Rottman,F.M.,Maroney,P.A.andNilsen,T.W.(1987)Mol.Cell.Biol.7, 1572-1575.
6.Davis,R.E.,Davis,A.H.,Carroll,S.M.,Rajkovic,A.andRottman,F.M.(1988)Mol.Cell Biol.8,4745-4755.
7.Donis-Keller,H.(1979)Nucl.Acids Res.7,179-192.
8.Goodwin,E.C.andRottman,F.M.(1992)Nucl. Acids Res.20,916.
RNase H
81For bulk or alternate pack sizes, email us at [email protected].
Molecular Biology Enzymes | Nucleases—Non-Specific
70196Y 15,000 unitsCustoms by request
Source:Staphylococcus aureus
Description:MicrococcalNucleaseisasingle-anddouble-strand-edDNAandRNAendonucleasewhichyields3'nucleotides(1).Itismorespecificforsingle-strandednucleicacidsandshowspreferenceforAUorAT-richregions.Completedigestionyieldsmono-anddinucleotides.
Applications:�� RemovalofRNAandDNAfromproteinpreparations�� RemovalofmRNAinareticulocytelysate�� Analysisofchromatinstructure
Form:Lyophilizedpowder
Properties:IsoelectricPoint:9.6OptimumpH:8.8-9.2OptimumTemperature:37°CMolecularWeight:16.81kDa(2)
RequirementforDivalentCation:Ca2+foractivityInhibitors:EDTAorEGTA
Purity:Testedforcontaminatingproteases.
Solubility and storage conditions:1.Shorttermstorage:Dissolve1mg/mlMicrococcal
NucleaseindH2O.Storeat4°Cfor1-2weeks.2.Longtermstorage:DissolveindH2Ocontaining
20-50%glycerolat1mg/mlconcentration.Storeat-20°C.Donotfreezeandthawmorethanonetime.
Assay conditions: Thereactionmixturecontains28mMsodiumborate,pH8.0,1.4mMCaCl2,100µgDNA,andenzyme.Incubationisat37°Cfor30minutes.
Unit definition:OneunitistheamountofenzymewhichchangestheopticaldensityofaDNAsubstrateby1.0at260nmatpH8.0,37°C(3).
Concentration:≥5,000units/mg
Shipping and storage:Shippedondryice.Seepreviousinformation.
References:1.Alexander,M.,Heppel,L.A.andHurwitz,J.(1961)
J.Biol.Chem.236,3014-3019.2.Taniuchi,H.,Anfinsen,C.B.andSodja,A.(1967)
J.Biol.Chem.242,4752-4758.3.Heins,J.N.,Taniuchi,H.andAnfinsen,C.B.
(1966)Procedures in Nucleic Acid Research,eds.G.L.CantoniandD.R.Davies(HarperandRowPublishers,Inc.,NewYork),79.
4.Cone,J.L.,Cusumano,C.L.,Taniuchi,H.andAnfinsen,C.B.(1971)J.Biol.Chem.246,3103-3110.
5.Ausubel,F.,Brent,R.,Kingston,R.E.,Moore,D.D.,Seidman,J.G.,Smith,J.A.andStruhl,K.(1989)Current Protocols in Molecular Biology.(JohnWileyandSons).
Micrococcal Nuclease (E.C.3.1.31.1)
20240Y 100 unitsCustoms by request
Source:Crotalus adamanteusvenom
Description:PhosphodiesteraseI(venomexonuclease)iswidelyusedinstudyingnucleicacidstructureandsequence(1).Ithydrolyzes5'-mononucleotidesfrom3'-hydroxy-terminatedDNAandRNA(2).Phosphodi-esteraseIcleavesADP-ribosylatedproteinsatthepy-rophosphatelinkagestoyieldphosphoribosyl-AMP(3).
Application:�� Basecompositionand“nearestneighbor”analysis(1)
Form:Lyophilizedpowder
Properties:MolecularWeight:115kDaOptimumpH:9.8-10.4Activators:Mg2+
Inhibitors:5mMEDTA,reducingagents(gluthathione,cysteine,andascorbicacid),partialinhibitionbyATP,ADP,andAMP
Purity:PartiallypurifiedbythemethodofBjörk(4)andtreatedtoremovecontaminating5'nucleotidase(5).
Assay conditions: Thereactionmixturecontains100mMTris-HCl,pH8.9,100mMNaCl,14mMMgCl2,0.5mMp-nitrophenylthymidine-5'-phosphate,andPhospho-diesteraseI.
Unit definition:Oneunithydrolyzes1µmolp-nitrophenylthymidine-5-phosphateperminutesat25°C,pH8.9.
Concentration:≥20units/mg
Resuspension and Storage: Dissolveat1mg/mlin110mMTris-HCl,pH8.9,110mMNaCl,15mMMgCl2,and50%glycerol.Storeat-20°Cafterrehydration.
Shipping and storage:Shippedondryice.Storeat-20°C.
References:1.Ho,N.W.andGilham,P.T.(1973)Biochem.
Biophys.Acta,308,53-58.2.Laskowski,M.(1971)inThe Enzymes, 3rd Ed.
(Academic Press, New York)IV,313-328.3.Hayaishi,O.(1976)Trends Biochem.Sci.1,9.4.Björk,W.,(1963)J. Biol. Chem.238,2487.5.Sulkowski,E.andLaskowski,M.(1971)Biochem.
Biophys.Acta,240,443.
Phosphodiesterase I(E.C.3.1.4.1)
82 888-362-2447 | 216-765-5000 | usb.affymetrix.com
Molecular Biology Enzymes | Phosphatases—Alkaline Phosphatases
70034Y 1,500 unitsCustoms by request
Source:Bovineintestinalmucosa
Description:CalfIntestinalAlkalinePhosphatase(CIAP)catalyzesthehydrolysisofmanyphosphomonoesters,includ-ing5'-nucleotides,RNA,andDNA.Itiswidelyusedforthedephosphorylationof5'-phosphorylatedendsofDNAorRNAforsubsequentlabelingwith32Pusing[g-32P]ATPandT4PolynucleotideKinase(PN70031).Dephosphorylationalsopreventsreliga-tionoflinearizedplasmidDNAincloningexperi-ments.CIAPcanbeinactivatedbyphenol:chloroformextraction,bygelpurification,orbyspincolumn.
Applications:�� Dephosphorylationof5'terminiofnucleicacidsbeforelabelingwithT4PolynucleotideKinase�� Dephosphorylationof5'terminiofDNAfragmentstopreventself-ligation
Properties:MolecularWeight:Dimerofapproximately110kDa
(two55kDasubunits)Activators:Zn2+,Mg2+,Ca2+
IsoelectricPoint:5.7RequirementforStability:TritonX-100,zincacetate
orzincchloride,cysteineOptimumpH:9-10OptimumTemperature:37°C.pHStability:pH7.5;5.0inthepresenceofTriton
X-100,Zn2+,cysteine
Purity:Testedforcontaminatingnon-specificendonucle-ases,exonucleases,andribonucleases.
Storage buffer:10mMTris-HCl(pH7.5),5mMMgCl2,0.1mMZnCl2,50%glycerol.
Assay conditions:Thereactionmixturecontains1Mdiethanolamine,pH9.8,0.25mMMgCl2,10mMp-nitrophenylphosphate,and0.01-0.1unitsofCalfIntestinalAlkalinePhosphatase(CIAP).Reactiontemperatureis37°C.Thechangeinabsorbanceat405nmismonitored(3050µlreactionvolume).
Unit definition:Oneunitofenzymecatalyzesthehydrolysisof1µmolofp-nitrophenylphosphateperminuteatpH9.8indiethanolaminebufferat37°C.
Concentration:20units/µl
Functional test:Dephosphorylationofarestrictionenzymedigestedplasmid(5-10pmolof5'ends,0.01-0.05units/pmol5'ends.Reducesreligationto<0.5%com-paredtotheuntreatedcontrol.)
Functionally tested 10X CIAP Reaction Buffer (1 ml included):200mMTris-HCl(pH8.0),100mMMgCl2.
CIAP Dilution Buffer, 10X (1 ml included):50mMTris-HCl(pH8.0).
Shipping and storage:Shippedondryice.Storeat-20°C.
References:1.Zhang,Y.,Zhu,Y.andZhou,H.(2000)Int. J.
Biochem. Cell. Biol.32,887-894.2.Abdolrazaghi,Z.andButterworth,P.(1983)
Enzyme30,12-20.3.Baunoch,D.A.,et al.(1992)Oncogene,7,2351-
2353.
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Shrimp Alkaline Phosphatase78390 500 units 1,000 units 5,000 units
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Calf Intestinal Alkaline Phosphatase (CIAP) (E.C.3.1.3.1.)
83For bulk or alternate pack sizes, email us at [email protected].
78390 500 units 1,000 units 5,000 units
Proven performance – the phosphatase benchmark�� Purifiedfromarecombinantsource�� 100%heat-inactivatedin5minutesat65°C�� Significantlyimprovedstoragestabilityatlowertemperatures(seeFig.1)�� Veryhighspecificactivity�� Removes5’-phosphatesfromDNA,RNA,dNTPs,andproteins�� Maybeaddeddirectlytorestrictionenzymedigests�� Novectorpurificationnecessary�� Requiresnosupplementalzincorotheradditivesforactivity�� Worksdirectinmanydifferentbuffers�� EasytreatmentofunincorporateddNTPsinPCRproductspriortoDNAsequencingorSNPanalysis
Source: Recombinant
Description:ShrimpAlkalinePhosphatase(SAP)isahighspecificactivity,heat-labilealkalinephosphatasepurifiedfromarecombinantsourceandoriginallyisolatedfromPandalus borealis(arcticshrimp).SAPisusefulinmanymolecularbiologyapplicationssuchasthedephosphorylationofphosphorylatedendsofDNAorRNAforsubsequentuseincloningorend-label-ingofprobes.Incloning,dephosphorylationpreventsreligationoflinearizedplasmidDNA.SAPmayalsobeusedtotreatunincorporateddNTPsinPCRreac-tionstopreparetemplatesforDNAsequencingorSNPanalysis.
SAPhasapproximatelythesamespecificactivityasCalfIntestinalAlkalinePhosphatase(CIAP),andlikeCIAP,isactiveinvirtuallyallrestrictionenzymereactionbuffers.UnlikeCIAP,SAPiscompletelyandirreversiblyinactivatedbyheatingreactionsat65°Cfor15minutes.
Applications:�� DephosphorylationofDNApriortocloning�� PCRcleanupforsequencingandSNPapplications�� DephosphorylationofDNApriortoend-labellingusingT4PNKorOptiKinase�� DephosphorylationofRNA�� Proteindephosphorylation
Properties:MolecularWeight:Homodimer.Monomeris55kDa
asdeterminedbyaminoacidsequence.OptimumpH:10.4inglycinebufferandpH8.0in
Trisbuffer.OptimumTemperature:37°CHeat-Inactivation:65°Cfor15minutesInhibitors:10mMDTT,0.1%β-MEReactionConditions:ActiveinNaClandKCl.
RequiresMg2+forhighestactivity.
Purity:Testedforcontaminatingendonucleases,exonucle-ases,andribonucleases.
Storage buffer:25mMTris-HCl(pH7.5),1mMMgCl2,50%glycerol.
Assay conditions:Thereactionmixturecontains100mMglycine,pH10.4,1mMMgCl2,1mMZnCl2,10mMp-nitrophenylphosphate,and0.001–0.1unitsofShrimpAlkalinePhosphatase(SAP).Thechangeinabsorbanceat405nmismonitored(3050μlreac-tionvolume).
Unit definition:Oneunitistheamountofenzymewhichcatalyzesthehydrolysisof1μmolofp-nitrophenylphosphateperminuteinglycinebuffer(pH10.4)at37°C.
Concentration:1unit/μl
Functional test:Dephosphorylationofrestrictionenzymedigestedplasmids(5–20pmolof5’-ends,0.1–0.5units/pmol5’-ends).Reducesreligationto<0.5%com-paredtotheuntreatedcontrol.
Functionally tested 10X SAP Reaction Buffer (1 ml included):200mMTris-HCl(pH8.0),100mMMgCl2.
Functionally tested SAP Dilution Buffer (1 ml included):50mMTris-HCl(pH8.0).
References:1.Ruan,C.C.,Samols,S.B.andFuller,C.W.(1990)
Comments17,(No.1),UnitedStatesBiochemicalCorporation,Cleveland,OH.
2.Werle,E.,ScneiderC.,Renner,M.,Völker,M.andFiehn,W.(1994)Nucl. Acids Res.22,4354-4355.
3.Hanke,M.andWink,M.(1994)BioTechniques17,858-860.
Shrimp Alkaline Phosphatase (SAP)
Molecular Biology Enzymes | Phosphatases—Alkaline Phosphatases
Fig. 1. Exceptional stability of SAP. ThreemonthstabilityofSAPatvarioustemperatures.SAPwasstoredfor3monthsattheindicat-edtemperaturesandtheremainingactivitywasmeasuredusingastandardp-nitrophenylphosphataseactivity.
0%
20%
40%
60%
80%
100%
-20ºC 4ºC 25ºC 37ºC
Rem
aini
ng A
ctiv
ity
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84 888-362-2447 | 216-765-5000 | usb.affymetrix.com
Molecular Biology Enzymes | Phosphatases—Alkaline Phosphatases
75592 50 reactions
Source:RecombinantPandalus borealis(arcticshrimp)
Description:SuperSAPisanenhancedformulationofShrimpAlkalinePhosphatasedesignedforrapiddephos-phorylationofDNApriortoitsuseincloningapplicationsandend-labeling.SuperSAPdephos-phorylatesupto5pmolsofcohesiveorbluntendswithabackgroundreduction≥95%injust5minutes.Backgroundreductionistheinhibitionofrecircularizationinaself-ligationreactionandmeasuredbytransformationinE. coli.SuperSAPmayalsobeusedtodephosphorylateRNAandprotein.
BenefitsSuperSAPhasallthebenefitsofSAP.Ithasahighspecificactivityandisheat-labile.SuperSAPcanbeeasilyheat-inactivatedat65°Cfor15minutesandworkswellinvariousrestrictionenzymebuffers.Noadditionalbuffersoradditivesarerequired.
Rapid dephosphorylationSuperSAPremovestheterminal5'phosphatefromprotruding-,blunt-orrecessed-endsinjust5minutes(Table1).
Easy vector preparation for cloningSuperSAPworksdirectlyinmostrestrictionenzymedigestsandcanbeaddeddirectlytothereactiontosimultaneouslycutanddephosphorylateDNA.Noadditionalbuffersoradditivesarerequired(Table2).
Convenient – Direct ligationSuperSAPreactionsdonotrequirepurification,e.g.phenolextraction,ethanolprecipitation,orspincol-umn,priortoligation.AfterSuperSAPtreatment,thedephosphorylatedcutvectormaybedirectlyligatedusingtheLigate-ITRapidLigationKit[PN78400/10]orT4DNALigase[PN70005].
Purity:Testedforcontaminatingendonucleases,exonucle-ases,andribonucleases.
Storage buffer:25mMTris-HCl,pH7.6,1.0mMMgCl2,0.1mMZnCl2,and50%glycerol.
10X SuperSAP Reaction Buffer (1 ml included): 200mMTris-HCl,pH8.0,100mMMgCl2.
Functional test:SuperSAPhasbeenfunctionallytestedinthefollow-ingprotocols:
Functional test: 5 Minute DephosphorylationDephosphorylationof5pmolofpurifiedvectorDNAterminiwith1µlSuperSAPin1XReactionBufferin5minutesat37°Creducesthebackgroundofreligationandtransformationto>95%comparedtountreatedcontrol.
Simultaneous dephosphorylation and restric-tion enzyme digestionSimultaneousdephosphorylationandrestrictionenzymedigestwith1µlofSuperSAPfor5'overhangandbluntendsand2µlfor3'overhangin30min-utesat37°Creducesthebackgroundofreligationandtransformationto>95%comparedtountreatedcontrol.
Components: 50Reaction PackSizeSuperSAP 50µl10XSuperSAPReactionBuffer 1ml
Shipping and storage:Shippedondryice.Storeat-20°C.
References:1.Ruan,C.C.,Samols,S.B.,andFuller,C.W.(1990)
Comments17,(No.1),UnitedStatesBiochemicalCorporation,Cleveland,OH.
2.Werle,E.,Scneider,C.,Renner,M.,Voelker,M.andFiehn,W.(1994)Nucl. Acids Res.22,4354-4355.
3.Hanke,M.andWink,M.(1994)BioTechniques17,858-860.
Table 1
Number of Background colonies reduction (%)
5' OverhangEcoR I-cut pUC 19 vectorControl 5831µlSuperSAP 5 99%
3' OverhangPst I-cut pUC 19 vectorControl 19871µlSuperSAP 0 100%
Blunt EndHinc II-cut pUC 19 vector Control 4901µlSuperSAP 2 99.6%
Rapid SuperSAP dephosphorylation for 5 minutes prevents recircularization of vector. pUC19vectorwasdigestedasindicat-ed,purifiedusingaspincolumnandre-suspendedin10mMTris-HCl,pH8.5.5µgwastreatedwith1µlofSuperSAPandincubatedfor5minutesat37°Cfollowedbyheat-inactivationat65°Cfor15min-utes.50ngwasself-ligateddirectlyusingtheLigate-ITRapidLigationKit(PN78400),2.5ngwastransformedintoE. coliDH5-aand0.5ngwasplatedonselectivemedium.
Table 2
Number of Background colonies reduction (%)
Kpn I-cut pUC 19 vector Buffer L (USB) 3' Overhang Control 47962µlSuperSAP 44 99%
Hind III-cut pUC 19 vector Buffer M (USB) 5' Overhang Control 17831µlSuperSAP 13 99.3%
EcoR I-cut pUC 19 vector Buffer H (USB) 5' Overhang Control 32691µlSuperSAP 84 97.4%
BamH I-cut pUC 19 vector Buffer K (USB) 5' Overhang Control 17531µlSuperSAP 0 100%
Sma I-cut pUC 19 vector Buffer A (USB) Blunt End Control 10911µlSuperSAP 0 100%
Simultaneous dephosphorylation with SuperSAP and restriction enzyme digestion is convenient and effective to prevent recircularization of vector. 1µgofpUC19wasdigestedasindicatedwith1µlofSuperSAP(5'overhangorblunt),or2µlofSuperSAP(3'overhang)intheappropriaterestrictionenzymebufferandwasincubatedfor30minutesat37°Cfollowedbyheat-inactiva-tionat65°Cfor15minutes.50ngwasdirectlyself-ligatedusingtheLigate-ITRapidLigationKit(PN78400),2.5ngwastransformedintoE. coliDH5-aand0.5ngwasplatedonselectivemedium.
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Molecular Biology Enzymes | Phosphatases—Pyrophosphatase
78450 5 units 50 unitsCustoms by request
Source:PurifiedfromE. colicellsexpressingtheclonedppagene(1)fromSaccharomyces cerevisiae(Baker’sYeast).
Description:DNAandRNApolymerasereactions,aswellasotherenzymaticreactions,generateinorganicpyrophos-phateasaby-product.Thisby-productinhibitsnucleicacidsynthesisthroughaprocesscalledpy-rophosphorolysiswhichreversesthepolymerizationreactionandlimitsDNA/RNAsynthesis.InorganicpyrophosphataseispartoftheDNA/RNAsynthesispathwayin vivo,renderingpolymerizationessentiallyirreversiblebyremovingpyrophosphate(2).Inorganicpyrophosphatase(PPase)catalyzesthehydrolysisofinorganicpyrophosphatetoformtwomoleculesoforthophosphate(3):P2O7
-4+H2O→2HPO4-2
Inorganicpyrophosphatasemaybeusedtoremovepyrophosphateandimprovetheresultsoftechniquesthatareparticularlysensitivetoitsaccumulationsuchassingle-baseextension(SBE)(4)andSangersequencing,especiallywithregardtoselectivebandweakening(5,6).Also,inorganicpyrophosphatasecangreatlyincreasetheyieldofin vitrotranscriptionreactions(7).
Applications:�� PreventsaccumulationofpyrophosphateduringDNAsequencing.�� High-yieldsynthesisofRNAbyin vitrotranscription.�� RemovalofcontaminantPPipriortoSNPgenotypingbasedonthedetectionofpyrophosphate.�� RemovalofcontaminantPPipriortosingle-base-extension(SBE).
Properties:MolecularWeight:70kDahomodimer(8)
(2x35kDa)pI:4.75Activators:Mg+2>Zn+2>Co+2>Mn+2>Ca+2,note
thathydrolysisofinorganicpyrophosphateisspecificinthepresenceofMg+2butbothADPandATPcanbehydrolyzedifzincispresent(9).
Inhibitors:EDTAInactivation:Inactivatedbyphenol/chloroform
extractionIVT:RecommendedconcentrationofrPPaseduringin
vitrotranscriptionreactionsis1-10units/ml
Purity:Greaterthan95%pureasdeterminedbySDS-PAGE.Testedforcontaminatingexonucleases,endonucle-ases,andribonucleases.
Storage buffer:10mMTris-HCl(pH7.5),0.1mMEDTA,50%glycerol.
rPPase Dilution Buffer (1 ml included):10mMTris-HCl(pH7.5),0.1mMEDTA,50%glycerol.
Unit definition:Oneunithydrolyzes1µmolofpyrophosphatein1minuteat25°C.
Concentration:0.1units/µl
Functional test:DNAsequencingusingtheSequenaseVersion2.0DNASequencingKit.
Shipping and storage:Shippedondryice.Storeat-20°C
References:1.Kurilova,S.A.,Vorobjeva,N.N.,Nazarova,T.I.
andAvaeva,S.M.(1993)FEBS Letters333(3),280-282.
2.Kornberg,A.(1980)DNAReplication(W.H.Freeman&Co.SanFrancisco),54-55.
3.Cooperman,B.S.(1982)Methods in Enzymology87,526-548.
4.Zhou,G.H.,Shirakura,H.,Kamahori,M.,OkanoK.,NagaiK.andKambara,H.(2004)Human Mutation24(2),155-163.
5.Tabor,S.andRichardson,C.C.(1990)J. Biol. Chem.265,8322-8328.
6.Ruan,C.C.,Samols,S.B.andFuller,C.W.(1990)Comments17,No.2,UnitedStatesBiochemicalCorporation,Cleveland,OH.
7.Cunningham,P.R.andOfengand,J.(1990)BioTechniques9,713-714.
8.Butler,L.G.(1971)Enzymes(AcademicPressNewYork),529-540.
9.Knight,W.B.,Dunaway-Mariano,D.,Ransom,S.C.andVillafranca,J.J.(1984)J. Biol. Chem.259(5),2886-2895.
Related product
Sequenase Version 2.0 DNA Polymerase with Pyrophosphatase70175 325 units
Pyrophosphatase, Inorganic (Recombinant) (rPPase)
86 888-362-2447 | 216-765-5000 | usb.affymetrix.com
70057Y 125 units70057Z 750 units70057 2,500 unitsCustoms by request
Source:E. coli straincontaininganoverproducingcloneofE. coli Exonuclease-freeKlenowfragment.
Description:ThelargefragmentofDNApolymeraseIfromE. coli,Klenow,hasbeengeneticallyengineeredtoeliminatethe3'→5'exonucleaseactivitynormallyassociatedwiththispolymerase(1).Exo-freeKlenowisusefulinapplicationswhereaccompanyingexonucleaseactivityisofsignificantconcern,suchasrandomprimedlabeling,DNAsequencingbytheSanger,et al.method,orstranddisplacementamplification(SDA).
Applications:�� RandomprimedDNAlabeling�� Stranddisplacementamplification(SDA)(2)
�� DNAsequencingbytheSangerdideoxymethod(3)
�� Labelingrecessed3'terminiofDNAfragmentswithlabeleddNTP
Note:Exonuclease-FreeKlenowisnotrecommendedforfill-inreactionsbeforeDNAligation,sinceitfrequentlyaddsoneormoreextranucleotidestothe3'-terminusofablunt-endedDNAsubstrateinanon-templatedirectedfashion(4).
Properties:MolecularWeight:68kDa
Purity:Greaterthan95%pureasdeterminedbySDS-PAGE.Testedforcontaminatingdouble-strandedendonu-cleasesandexonucleases.
Storage buffer:50mMpotassiumphosphate(pH7.0),1.0mMDTT,1.0mMEDTA,50%glycerol.
Assay conditions: Thereactionmixturecontains100mMpotassiumphosphate(pH7.4),6.7mMMgCl2,1mMDTT,100µMeachdNTP,radiolabeleddATP,400µg/mlactivatedsalmonspermDNA,andenzyme.Incuba-tionisfor30minutesat37°C.
Unit definition:Oneunitistheamountofenzymerequiredtocatalyzetheincorporationof10nmoloftotaldeoxy-ribonucleotidesintoanacid-precipitablematerialin30minutesat37°C.
Concentration:10units/µl
Functional test:Filling-in3'recessedendswith>50%incorporationofradiolabeleddATPinto4µgofrestrictionenzymedigestedDNAin15minutesat30°C.
Functionally tested 10X Exonuclease-Free Klenow Reaction Buffer (1 ml included):0.5MTris-HCl(pH7.4),0.1MMgCl2,10mMDTT.
Shipping and storage:Shippedondryice.Storeat-20°C.
References:1.Derbyshire,V.,Freemont,P.S.,Sanderson,M.R.,
Beese,L.,Friedman,J.M.,Joyce,C.M.andSteitz,T.A.(1988)Science 240,199-201.
2.Walker,G.T.(1993)PCR Methods Appl.3,1-6.3.Sanger,F.,Nicklen,S.andCoulsen,A.R.(1977)
Proc. Natl. Acad. Sci. U.S.A. 74,5463-5467.4.Clark,J.M.,Joyce,C.M.andBeardsley,G.P.
(1987)J. Mol. Biol.198,123-127.
Exonuclease-Free Klenow (LargeFragmentofDNAPolymeraseI,Exonuclease-Free,E.C.2.7.7.7)
Molecular Biology Enzymes | Polymerases—DNA Polymerases
72020 750 units 2,500 units
Source:E. colistraincontaininganoverproducingcloneofE. coliExonuclease-freeKlenowfragment.
Description:ThelargefragmentofDNApolymeraseIfromE. coli,Klenow,hasbeengeneticallyengineeredtoeliminatethe3’→5’exonucleaseactivitynormallyassociatedwiththispolymerase(1).Exo-freeKlenowisusefulinapplicationswhereaccompanyingexonucleaseactivityisofsignificantconcern,suchasrandomprimedlabeling,DNAsequencingbytheSanger,et al.method,stranddisplacementamplifica-tion(SDA),orwholegenomeamplification(WGA).Exonuclease-FreeKlenow,TrisBufferisstoredinaphosphatefreebuffer
Applications:�� RandomprimedDNAlabeling�� Stranddisplacementamplification(SDA)(2)
�� DNAsequencingbytheSangerdideoxymethod(3)
�� Labelingrecessed3'terminiofDNAfragmentswithlabeleddNTP
Note:Exonuclease-FreeKlenowisnotrecommendedforfill-inreactionsbeforeDNAligation,sinceitfrequentlyaddsoneormoreextranucleotidestothe3'-terminusofablunt-endedDNAsubstrateinanon-templatedirectedfashion(4).
Properties:MolecularWeight:68kDa
Purity:Greaterthan95%pureasdeterminedbySDS-PAGE.Testedforcontaminatingdouble-andsingle-strand-edendonucleasesandexonucleases.
Storage buffer:25mMTris-HCl,pH7.5,1mMDTT,0.1mMEDTAand50%glycerol
Assay conditions: Thereactionmixturecontains100mMpotassiumphosphate(pH7.4),6.7mMMgCl2,1mMDTT,20μMpoly(dA-dT)-(dA-dT),33μMdATP,33μMdTTP,andenzyme.Incubationisfor30minutesat37°C.
Unit definition:Oneunitistheamountofenzymerequiredtocatalyzetheincorporationof10nmoloftotaldeoxy-ribonucleotidesintoanacid-precipitablematerialin30minutesat37°C.
Concentration:10units/µl
Functional test:Filling-in3’recessedendswith>50%incorporationofradiolabeleddATPinto0.1-4μgofrestrictedDNAin15minutesat30°C.
Functionally Tested 10X Exonuclease-Free Klenow Reaction Buffer (1 ml included):0.5MTris-HCl(pH7.4),0.1MMgCl2,10mMDTT.
References:1.Derbyshire,V.,Freemont,P.S.,Sanderson,M.R.,
Beese,L.,Friedman,J.M.,Joyce,C.M.andSteitz,T.A.(1988)Science 240,199-201.
2.Walker,G.T.(1993)PCR Methods Appl.3,1-6.3.Sanger,F.,Nicklen,S.andCoulsen,A.R.(1977)
Proc. Natl. Acad. Sci. U.S.A. 74,5463-5467.4.Clark,J.M.,Joyce,C.M.andBeardsley,G.P.
(1987)J. Mol. Biol.198,123-127.
Exonuclease-Free Klenow, Tris Buffer(LargeFragmentofDNAPolymeraseI,Exonuclease-Free)
87For bulk or alternate pack sizes, email us at [email protected].
Molecular Biology Enzymes | Polymerases—DNA Polymerases
2141Y 300 units2141Z 1,500 unitsCustoms by request
Source:E. coli straincontaininganoverproducingcloneofE. coli Klenowfragment.
Description:Klenowenzyme,thelargefragmentofDNAPolymeraseIfromE. coli,retainspolymerizationand3'→5'exonucleaseactivitybutlacksthe5'→3'exonucleaseactivity.Itiswell-suitedfortheconversionof5'-protrudingendstoflushendsforcloning(1),in3'-endlabelingofDNAfragmentsforsequenceanalysis(2),forsecond-strandsynthesisofcDNA(3),andforrandomprimedlabelingofDNAfragments.
Applications:�� Fillinof3'recessedterminiproducedbysomerestrictionenzymesbeforeligation�� SecondstrandcDNAsynthesis�� Labeling3'terminiofDNAfragmentswithlabeleddNTP
Properties:MolecularWeight:68kDaOptimumpH:7.4inphosphatebuffer;8.4inTris-HClOptimumTemperature:37°CRequirementsforDivalentCation:Mg2+,Mn2+
OptimumMg2+Concentration:7mM
Purity:Greaterthan98%pureasdeterminedbySDS-PAGE.Testedforcontaminatingdouble-andsingle-strandedendonucleases.
Storage buffer:50mMpotassiumphosphate(pH7.0),1.0mMDTT,50%glycerol.
Assay conditions:Thereactionmixturecontains100mMpotassiumphosphate(pH7.4),6.7mMMgCl2,1mMDTT,100µMeachdNTP,radiolabeleddATP,400µg/mlactivatedsalmonspermDNA,andenzyme.Incuba-tionisfor30minutesat37°C.
Unit definition:Oneunitistheamountofenzymerequiredtocatalyzetheincorporationof10nmoloftotaldeoxy-ribonucleotidesintoanacid-precipitablematerialin30minutesat37°C.
Concentration:5units/µl
Functional test:Filling-in3'recessedendswith>50%incorporationofradiolabeleddATPinto4µgofrestrictionenzymedigestedDNAin15minutesat30°C.
Functionally tested 10X Klenow Reaction Buf-fer (1 ml included):0.5MTris-HCl(pH7.5),100mMMgCl2,10mMDTT,0.5mg/mlBSA.
Shipping and storage:Shippedondryice.Storeat-20°C.
References:1.Telford,J.L.,Kressmann,A.,Koski,R.A.,
Grosschedl,R.,Muller,F.,Clarkson,S.G.andBirnstiel,M.L.(1979)Proc. Natl. Acad. Sci.USA 76,2590-2594.
2.Joyce,C.M.andGrindley,N.D.F.(1983)Proc. Natl. Acad. Sci. USA 80,1830-1834.
3.Rougeon,F.,Kourilsky,P.andMach,B.(1975)Nucl. Acids Res. 2,2365-2378.
Klenow DNA Polymerase I (LargeFragmentofDNAPolymeraseI,E.C.2.7.7.7)
70010Y 500 unitsCustoms by request
Source:E. coli straincontaininganoverproducingcloneofE. coli DNAPolymeraseI.
Description:DNAPolymeraseIisaDNAdependent5'→3'polymerasewithinherent3'→5'and5'→3'exonucleaseactivity.Thesethreeactivitiestogethermakethisenzymeusefulforin vitrolabelingofDNAbythenicktranslationmethod(1).ThismethodinvolvestheuseofalowlevelofDNaseIwhichhydrolyzesphosphodiesterbondstocreatenicksatinternallocationsinunlabeledDNA.The3'hydroxylgroupsatthenickingsiteprimeDNAsynthesisbyDNAPolymeraseIwhichconcomitantlydegradestheDNA5'ofthenick.Thisprocess(translation)leavesaradiolabeledpatchofDNAwhichextendsfromtheoriginalnicksitetothesiteoftheremainingnewnickorterminusofafragment.
Applications:�� NicktranslationofDNAinmakingprobes�� Second-strandsynthesisofcDNA
Properties:MolecularWeight:109kDaOptimumpH:7.4inphosphatebuffer;8.4inTris-HClbufferOptimumTemperature:37°CRequirementsforDivalentCation:Mg2+,Mn2+
OptimumMg2+Concentration:7mM
Purity:Greaterthan90%pureasdeterminedbySDS-PAGE.Testedforcontaminatingdouble-strandedendonucleases.
Storage buffer:50mMpotassiumphosphate(pH7.0),1.0mMDTT,50%glycerol,1mMEDTA.
Assay conditions:Thereactionmixturecontains100mMpotas-siumphosphate(pH7.4),6.7mMMgCl2,1.0mMDTT,150µMdNTPs,400µg/mlactivatedDNAastemplate-primer,andenzyme.Incubationisat37°Cfor30minutes.
Unit definition:Oneunitistheamountofenzymerequiredtocata-lyzetheconversionof10nmoloftotaldeoxyribonu-cleotidesintoanacid-insolubleformin30minutesat37°CusingactivatedDNAasthetemplate-primer.
Concentration:10units/µl
Shipping and storage:Shippedondryice.Storeat-20°C.
Reference:1.Rigby,P.W.,Dieckmann,M.,Rhodes,C.andBerg,
P.(1977)J. Mol. Biol.113,237-257.
DNA Polymerase I (E.C.2.7.7.7)
88 888-362-2447 | 216-765-5000 | usb.affymetrix.com
70775Y 200 units70775Z 1,000 unitsCustoms by request
Source:E. coli straincontainingclonesthatoverproduceT7Gene5ProteinandE. coli Thioredoxin.
Description:SequenaseVersion2.0DNAPolymeraseisageneticallyengineeredformofT7DNApolymerase(1).Unlikethewild-typeenzymeithasvirtuallyno3'→5'exonucleaseactivity.SequenaseVersion2.0ishighlyprocessive,incorporatesnucleotideanalogs(dlTP,thio-dNTPs,dideoxy-NTPs,etc.),isnotimpededbysecondarystructures,andcancarryoutstranddisplacementsynthesis.Itisanexcellentenzymefordideoxy-sequencingandisusefulinotherapplica-tions,especiallywheretheabsenceofassociatedexonucleaseactivityisdesirable.
Applications:�� DNAsequencing�� Secondstrandsynthesisformicroarrayapplications(2)
�� Randomprimedlabelingwithnucleotideanalogs(2)
�� Labelingthe3'terminiofDNAfragmentswith5'protrudingends�� AmplificationofDNA�� Stranddisplacementapplications
Note:SequenaseVersion2.0DNAPolymerasemaynotbeusefulformakingblunt-endfragmentsbecauseitcanleaveaone-baseover-hangatthe3'end.
Properties:MolecularWeight:Consistsoftwosubunits,
modifiedT7gene5protein(76kDa)andE. colithioredoxin(12kDa).
OptimumpH:7.5OptimumTemperature:37°CRequirementsforDivalentCation:Mg2+,Mn2+
Purity:Greaterthan95%pureasdeterminedbySDS-PAGE.Testedforcontaminatingdouble-andsingle-strand-edendonucleasesandexonucleases.
Storage buffer:20mMpotassiumphosphate(pH7.4),1mMDTT,0.1mMEDTA,50%glycerol.
Assay conditions:Thereactionmixture(100µl)contains40mMTris-HCl(pH7.5),10mMMgCl2,5mMDTT,0.3mMdNTPs,and5µgM13mp18pre-annealedto5pmolM13universalprimer;incubationisat37°Cfor1minute.
Unit definition:Oneunitofenzymecatalyzestheincorporationof1nmolofnucleotideintoacidinsolubleformin30secondsat37°C.
Concentration:13units/µl
Functional test:DNAsequencingwiththeSequenaseVersion2.0DNASequencingKit(PN70770).
Functionally tested 5X Sequenase Version 2.0 Reaction Buffer (1 ml included, PN 70702):200mMTris-HCl(pH7.5),100mMMgCl2,250mMNaCl
Sequenase Version 2.0 Dilution Buffer (1 ml included):10mMTris-HCl(pH7.5),5mMDTT,0.1mMEDTA
Shipping and storage:Shippedondryice.Storeat-20°C.
References:1.Tabor,S.andRichardson,C.C.(1989)J. Biol.
Chem.264,6447-6458.2.Wang,D.,Coscoy,L.,Zylberberg,M.,Avila,P.C.,
Boushey,H.A.,Ganem,D.andDeRisi,J.L.(2002)Proc Natl. Acad. Sci. USA,99,15687-15692.
3.Paris,M.(1992)Comments 18,(No.3),UnitedStatesBiochemicalCorporation,Cleveland,Ohio.
4.Tabor,S.andRichardson,C.C.(1987)Proc. Natl. Acad Sci. USA 84,4767-4771.
5.Tabor,S.andRichardson,C.C.(1989)Proc. Natl. Acad. Sci. USA 86,4076-4080.
Sequenase Version 2.0 DNA Polymerase (E.C.2.7.7.7)
Molecular Biology Enzymes | Polymerases—DNA Polymerases
70017Z 250 units70017 1,000 unitsCustoms by request
Source:E. coli straincontainingoverproducingclonesofT7Gene5andE. colithioredoxin.
Description:T7DNAPolymeraseconsistsoftwosubunits,oneencodedbythebacteriophagegene5andtheotherbytheE. coli trxAgenethioredoxin(1,2).Thisenzymeisresponsiblefortherapidreplicationofbacterio-phageT7DNAduringitsinfectioncycle.InadditiontothehighlyprocessiveDNApolymeraseactivity,italsohashighlevelsofbothsingle-anddouble-strandedDNA3'→5'exonucleaseactivities(3,4).Theexonucleasespecificactivitieshavebeenreportedtorangefrom5%to100%ofthepolymerasespecificactivity(3,5).Theexonucleaseactivityappearstoberesponsibleforthehighfidelityofthisenzymeandpreventsstranddisplacementsynthesis(6).
T7DNAPolymeraseisavaluableenzymewherehighexonucleaseactivityisrequiredorwhereabsenceofstrand-displacementactivityisimportant(6).Ithasalsobeenusedsuccessfullyforsite-directedmutagen-esis(7).This enzyme is not suitable for use in DNA sequencing.ItisdistinctfromSequenaseVersion2.0DNAPolymerasewhichisamodifiedformofT7DNAPolymerasewithno3'→5'exonucleaseactivity.
Applications:�� Labeling3'-terminiofDNAfragmentswithprotruding5'ends(Fill-InReaction)�� Labelingthe3'-terminiofblunt-endedDNAfragmentsortheterminiofDNAfragmentswithprotruding3'-termini�� Site-directedmutagenesis�� NotsuitableforDNAsequencingduetohighexogenouslevelsofexonucleaseactivities
Properties:T7Gene5product(84kDa)andE. colithioredoxin(12kDa).
Purity:Greaterthan95%pureasdeterminedbySDS-PAGE.Testedforcontaminatingendonucleases.
Storage buffer:20mMpotassiumphosphate(pH7.4),1mMDTT,0.1mMEDTA,50%glycerol.
Assay conditions:Thereactionmixturecontains88mMpotassiumphosphate(pH7.5),6.7mMMgCl2,5mM2-ME,300µMofeachdNTP,5mMactivatedDNA,andenzyme.
Unit definition:Oneunitistheamountofenzymerequiredtocata-lyzetheincorporationof10nmoloftotalnucleotideintoacidinsolubleformin30minutesat37°Cunderstandardassayconditions.
Concentration:10units/µl
5X T7 DNA Polymerase Reaction Buffer (1 ml included, PN 70097):200mMTris-HCl(pH7.5),100mMMgCl2,and250mMNaCl.
T7 DNA Polymerase Dilution Buffer (1 ml included):20mMpotassiumphosphatebuffer(pH7.4),1mMDTT,0.1mMEDTA,50%glycerol.
Shipping and storage:Shippedondryice.Storeat-20°C.
References:1.Grippo,P.andRichardson,C.C.(1971)J. Biol.
Chem.246,6867-6873.2.Modrich,P.andRichardson,C.C.(1975)J. Biol.
Chem.250,5515-5522.3.Adler,S.andModrich,P.(1979)J. Biol. Chem.
254,11605-11614.4.Hori,K.,Mark,D.F.andRichardson,C.C.(1979)
J. Biol. Chem.254,11598-11604.5.Engler,M.J.,Lechner,R.L.andRichardson,C.C.
(1983)J.Biol.Chem.258,11165-11173.6.Lechner,R.L.andRichardson,C.C.(1983)
J. Biol. Chem.258,11185-11196.7.Bebenek,K.et al.(1989)Nucl. Acids Res. 17,5408.
T7 DNA Polymerase (E.C.2.7.7.7)
89For bulk or alternate pack sizes, email us at [email protected].
Molecular Biology Enzymes | Polymerases—DNA Polymerases
70052 200 units 1,000 units
Source:E. colistrainexpressingafulllengthunmodifiedcloneofTthDNAPolymerasefromThermus ther-mophilusstrainHB8.
Description:TthDNAPolymeraseisathermostableenzymederivedfromThermus thermophilus(1)whichhasbeenclonedandpurifiedfromE. coli.Theenzymepossessesahighlyprocessive5'→3'polymeraseactivitybutlacksa3'→5'exonucleaseactivity.TthDNAPolymeraseissuitableforPCRinthepres-enceofMg2+ions.ItalsohasanintrinsicreversetranscriptaseactivityinthepresenceofMn2+ions.ThereversetranscriptionactivityusingRNAasatemplatetopreparecDNAmaybeachievedathightemperatures,therebyeliminatingproblemsrelatedtosecondarystructure.TthDNAPolymeraseisverystablewithahalf-lifeof20minutesat95°C.
TthDNAPolymeraseissuitableforRT-PCRorforPCRalone.Theenzymeissuppliedwith5XRTBuffer,5XChelateBuffer,10XPCRBuffer,andseparatetubesof9mMMnCl2and25mMMgCl2.
Applications:�� Hightemperaturereversetranscriptionbetween60°-70°Ctohelpeliminatesecondarystructure�� QuantitationofviralRNAinclinicalsamples(2)
�� DetectionofmRNAinindividualcellsbyRT-PCR(3)
�� RapiddetectionofbacterialDNAsamplesbyPCRwithoutDNAextractionorcelllysis(4)
�� RT-PCRinthepresenceof2-5%(vol/vol)ofphenolsaturatedbuffer(5)
Properties:MolecularWeight:94kDaActivator:Mn2+forRNAdependentDNApolymerase
activity;Mg2+forDNAdependentDNApolymeraseactivity.
Purity:Greaterthan95%pureasdeterminedbySDS-PAGE.Freefromdetectablenon-specificendoribonucleases.
Storage buffer:20mMTris-HCl(pH8.5),1mMDTT,0.1mMEDTA,100mMKCI,50%glycerol,stabilizers.
Assay conditions:Thereactionmixturecontains67mMTris-HCl,pH8.8(at25°C),16.6mM(NH4)2SO4,6.7mMMgCl2,10mMβ-ME,200µMeachdATP,dGTP,dTTP,100µMdCTP,radio-labeleddCTP,250µg/mlactivat-edfishspermDNA,andTthDNAPolymerase.Afterincubationat74°Cfor10minutes,acidinsolublematerialisdetermined(50µlreactionvolume).
Unit definition:Oneunitincorporates10nmoloftotalnucleotidesintoacid-insolublematerialin30minutesat74°Cinatotalvolumeof50µl.
Concentration:5units/µl
Functional test:TthDNAPolymeraseisfunctionallytestedforprod-uctyieldandlengthinRT-PCRandPCRamplifica-tion.
5X RT Buffer (included): 50mMTris-HCl,pH8.9,450mMKCl.
5X Chelate Buffer (included): 50mMTris-HCl,pH8.9,500mMKCl,3.75mMEGTA,0.25%Tween20,7.5mMMgCl2,25%glycerol.
10X PCR Reaction Buffer (included): 100mMTris-HCl,pH8.6,500mMKCl,15mMMgCl2.
25 mM MgCl2 (included): MgCl2 solution.
9 mM MnCl2 (included): MnCl2solution.
Shipping and storage:Shippedondryice.Storeat-20°C.
References:1.Ruttmann,C.M.,Corotas,M.,Zalvidar,J.and
Vicuna,R.(1985)Eur. J. Biochem.149,41-46.2.Mulder,J.,McKinney,N.,Christopherson,C.,
Sninsky,J.,Greenfield,L.andKwok,S.(1994)J. of Clinical Microbiology32,292-300.
3.Chiocchia,G.andSmith,K.A.(1997)BioTechniques 22,312-318.
4.Lofstrom,C.,Knutsson,R.,Axelsson,C.E.andRadstrom,P.(2004)Appl. and Environmental Microbiology.70,69-75.
5.Katcher,H.AndSchwartz,I.(1994)BioTechniques 16,84-92.
Tth DNA Polymerase
Fig. 1. RT/PCR of target with high G+C content (77%) 0.387 kb Notch3 (lane1) and 1.0 kb single copy gene Numb (lane 2).
Fig. 2. PCR of DNA fragments using Tth DNA Polymerase ranging from 1 kb to 6 kb from Lambda DNA.
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90 888-362-2447 | 216-765-5000 | usb.affymetrix.com
Molecular Biology Enzymes | Polymerases—Reverse Transcriptases
70041Y 200 units70041Z 1,000 unitsCustoms by request
Source:PurifiedvirionofAvian myeloblastosisvirus.
Description:AMVReverseTranscriptasecatalyzesthepolymeriza-tionofDNAusingtemplateDNA,RNAorRNA:DNAhybrids(1).Itconsistsoftwopolypeptidechains,oneofwhichcontainsa5'→3'polymeraseactivityandtheotheranRNaseHactivity.ReversetranscriptaserequiresMg2+orMn2+andisusedforfirststrandsynthesisofcomplementaryDNA(cDNA)frommRNAtemplate(2).Underproperconditions,highyieldsoffull-lengthcDNAcanbeobtainedwithAMVReverseTranscriptase(3).AMVReverseTranscriptaseisalsousefulintheamplificationofRNA(4).
Applications:�� SynthesisofcDNAforPCR,cloning,andhybridizationprobes�� Filling-inandlabelingthe3'terminiofDNAwith5'protrudingends�� RNAsequencing�� AmplificationofRNA(4)
�� Primerextensionassays
Properties:MolecularWeight:157kDaInactivation:75°Cfor10minutesorbyadding2µl
of0.5MEDTAfora50µlreaction.
Purity:Testedforcontaminatingendonucleases,exonucle-asesandribonucleases.
Storage buffer:200mMpotassiumphosphate(pH7.2),2mMDTT,0.2%TritonX-100,50%glycerol.
Assay conditions:Thereactionmixture(100µl)contains50mMTris-HCI(pH8.3),6mMMgCl2,40mMKCI,0.5mMradiolabeledTTP,and0.4mMPoly(rA)n:oligo(dT)50.Incubationisat37°Cfor10minutes.
Unit definition:Oneunitistheamountofenzymerequiredtoincor-porate1nmolofradiolabelednucleotideintoacidinsolubleproductin10minutesat37°C.
Concentration:15units/µl
Functional test:RT-PCRamplificationongenerationof0.5kband1.5kbRT-PCRproducts.
Functionally tested 5X AMV RT Reaction Buffer (1 ml included):250mMTris-HCl(pH8.3),40mMMgCl2,250mMNaCl,5mMDTT
Shipping and storage:Shippedondryice.Storeat-20°C.Avoidfreeze-thawcycles,whichresultinlossofenzymeactivity.
References:1.Kacian,D.L.(1977)inMethods in Virology,eds.
K.MaramoroschandH.Koprowski6,143-184.2.Goodman,H.M.andMacDonald,R.J.(1979)
Methods Enzymol.68,75-90.3.Berger,S.L.,Wallace,D.M.,Puskas,R.S.and
Eschenfeldt,W.H.(1983)Biochemistry22,2365-2372.
4.PCRProtocols:AGuidetoMethodsandApplications(1990)eds.M.A.Innis,D.H.Gelfand,andJohnJ.Sni,(AcademicPress,NewYork),23.
AMV Reverse Transcriptase
78306 25,000 units 100,000 unitsCustoms by request
M-MLV Reaction Buffer, 5X(Included with each pack size of enzyme)71505 1 ml
Source:E. colistraincontaininganoverproducingcloneofM-MLVReverseTranscriptase.
Description:M-MLVReverseTranscriptasecatalyzesthepolymerizationofDNAusingtemplateDNA,RNA,orRNA:DNAhybrids(1).Full-lengthcopiesoflargemRNAs,>10kb,maybesynthesized.M-MLVReverseTranscriptasehasamuchlowerRNaseHactivitythanAMVReverseTranscriptase,resultinginhighyieldsoffulllengthcDNA(2).ThismakesM-MLVReverseTranscriptaseveryusefulincDNAsynthesisandRT-PCR.
ForaddedconvenienceandoptimalRT-PCR,trytheRT-PCRKits(PN78350and78355)andRT-PCRMasterMixes(PN71185and78370).
Applications:�� RT-PCR�� SynthesisoffirststrandcDNAforPCR,cloning,andhybridizationprobes�� Filling-inandlabelingthe3'terminiofDNAwith5'protrudingends�� AmplificationofRNA�� Primerextensionassays
Properties:MolecularWeight:71kDa(monomeric).Inhibitors:Polyamines,phosphate,pyrophosphates,
andlithiumchloride(3,4).Inactivation:75°Cfor10minutesorbyadding2µl
of0.5MEDTAfora50µlreaction.
Purity:Greaterthan90%pureasdeterminedbySDS-PAGE.Testedforcontaminatingendonucleases,exonucle-ases,andribonucleases.
Storage buffer:20mMTris-HCl(pH7.5),0.1MNaCl,0.1mMEDTA,1mMDTT,0.01%IgepalCA-630,50%glycerol.
Assay conditions:Thereactionmixture(25µl)contains50mMTris-HCI(pH8.3),40mMKCl,6mMMgCl2,1mMDTT,400µMpoly(rA)-oligo(dT)12-18,500µMradiolabeledTTP,and0.1-0.5unitsenzyme.Incubationisat37°Cfor10minutes.
Unit definition:Oneunitisthatamountofenzymerequiredtoincorporate1nmolofdeoxynucleotideintoDE-81filter-bindingmaterialin10minutesat37°Cusingpoly(rA)-oligo(dT)12-18astemplate-primer.
Concentration:200units/µl
Functional test:RT-PCRamplificationongenerationof0.5kband1.5kbRT-PCRproducts.
Functionally tested 5X M-MLV Reaction Buffer (1 ml included, PN 71505):250mMTris-HCl(pH8.3),375mMKCl,15mMMgCl2,50mMDTT.
Shipping and storage:Shippedondryice.Storeat-20°C.
References:1.Roth,M.J.,Tanese,N.andGoff,S.P.(1985)
J.Biol.Chem.260,9326-9335.2.Sambrook,J.andRussell,D.W.(2001)Molecular
Cloning: A Laboratory Manual,3rdEdition,ColdSpringHarbor,NewYork,8.48.
3.Gerard,G.F.andD’Alessio,J.M.(1993)Methods In Molecular Biology16, HumanaPress,NJ,73-93.
4.Sambrook,J.andRussell,D.W.(2001)Molecular Cloning: A Laboratory Manual,3rdEdition,ColdSpringHarbor,NewYork,A8.16.
M-MLV Reverse Transcriptase(E.C.2.7.7.49)
91For bulk or alternate pack sizes, email us at [email protected].
Molecular Biology Enzymes | Polymerases—RNA Polymerases
78040Y 100 unitsCustoms by request
Source:E. coliRNAPolymeraseHoloenzymeisisolatedfromtherifampicin-sensitivestrainBL21andissigmasaturated.
Description:E. coliRNAPolymeraseisaholoenzymeusedfortranscriptionofDNAandtheproductionoflabeledRNAforhybridizationprobes.Thepolymerasecon-sistsof5subunitsdesignateda,a,β',β,andσ.Theenzymeis100%saturatedwithsigmafactorwhichisrequiredforcorrectinitiationofRNAsynthesisonDNAtemplatescontainingbacterialandphagepromoters.
Application:�� SynthesisoftranscriptswhenitisnotpossibletotranscribeDNAfromavectorcontainingaphageRNApromoter
Properties:MolecularWeight:Approximately450kDa.Themolecularweightforthesubunitsare36.5(each),156,151,and70kDafora,β',β,andσsubunitsrespectively.
Purity:Testedforcontaminatingnon-specificexonucleases,endonucleases,andribonucleases.
Storage buffer:50mMTris-HCl(pH7.5),250mMNaCl,0.1mMEDTA,1.0mMDTT,50%glycerol.
Assay conditions:Thereactionmixture(250µl)contains40mMTris-HCI(pH7.5),150mMKCI,10mMMgCl2,0.01%TritonX-100,10mMDTT,0.02µg/µlDNAtemplate,0.5mMofeachrNTP,andenzyme.Incubationisat37°Cfor10minutes.
Unit definition:Oneunitistheamountofenzymerequiredtocata-lyzetheincorporationof1nmolofaribonucleosidetriphosphateintoRNAin10minutesat37°C.
Concentration:1unit/µl
Shipping and storage:Shippedondryice.Storeat-20°C.
References:1.Burgess,R.R.andTravers,A.A.(1970)Fed Proc.
29,1164-1169.2.Burgess,R.R.,Travers,A.A.,Dunn,J.J.andBautz,
E.K.(1969)Nature221,43-46.3.Fujiki,H.,Palm,P.,Zillig,W.,Calendar,R.and
Sunshine,M.(1976)Mol.Gen.Genet.145,19-22.
E. coli RNA Polymerase Holoenzyme(RNANucleotidylTransferase,E.C.2.7.7.6)
74225Y 10,000 units74225Z 25,000 unitsCustoms by request
Poly(A) Polymerase Reaction Buffer, 5X(Included with each pack size of enzyme)74226 1 ml
Source:E. colistraincontaininganoverproducingcloneofYeastPoly(A)Polymerase.
Description:Poly(A)Polymerasecatalyzestheadditionofadeno-sineresiduesontothe3'-endsofRNA(1,2).Itcanbeusedtoaddpoly(A)tailstoRNAinthefirststepofcloning(3).ThereactionrequiresMn2+orMg2+,ATPassubstrate,andanyRNAcontaining3'hydroxylter-miniasprimer.LongerRNAmoleculesaresomewhatbetterprimersthanshortoligomers(4).Substitutionofcordycepin-5'-triphosphate(3'-dATP)forATPresultsinadditionofasingle3'-dAresiduetotheendsoftheRNA,ausefultechniqueforlabelingRNAatthe3'-end(5)*.
Incomparativestudies,YeastPoly(A)PolymeraseworksmoreefficientlythanE. coliPoly(A)Polym-eraseforRNAoligonucleotide-labelingandpoly(A)tailing.Shorterincubationtimesarerequiredfortheyeastenzymeanditisfoundtolabelbothlongandshortsubstratesequallywell.Poly(A)PolymeraseisrecommendedoverT4RNALigasefor3'-endlabel-ingoflongRNAmolecules.
*Note:Variousmodifiednucleotidescanalsobeusedtolabelthe3'endofRNAusingYeastPoly(A)Polymerase(5).
Applications:�� Additionofpoly(A)tailstoRNA�� Labelingthe3'endsofRNA
Purity:Ribonucleasefree.
Storage buffer:20mMTris-HCl(pH8.0),50mMKCl,0.5mMDTT,50%glycerol.
Assay conditions:25mMTris-HCl(pH7.0),40mMKCl,0.5mMMnCl2,0.5mMEDTA,0.5mMDTT,0.2mg/mlBSA,10%glycerol,3.3µMradiolabeledATP,0.5mMATP,6.5µgpoly(A)(~100bases),poly(A)polymerase.Afterincubationat37°Cfor10minutes,acidinsolubleradioactivityisdetermined.
Unit definition:Oneunitistheamountofenzymewhichincorpo-rates1pmolAMPintoacid-insolublematerialat37°Cin1minute.
Concentration:600units/µl
Functional test:3'-endlabelingofaribonucleotidewithcordycepin-5'-triphosphate.
Functionally tested 5X Poly(A) Polymerase Reaction Buffer (1 ml included, PN 74226):100mMTris-HCl(pH7.0),3.0mMMnCl2,0.1mMEDTA,1mMDTT,500µg/mlacetylatedBSA,50%glycerol.
Shipping and storage:Shippedondryice.Storeat-20°C.
References:1.Sippel,A.E.(1973)Eur.J.Biochem.37,31-40.2.Edmonds,M.(1982)inThe Enzymes,3rdedition,
ed.P.D.Boyer(AcademicPress,NewYork)15,217-244.
3.Gething,M.J.,Bye,J.,Skehel,J.andWaterfield,M.(1980)Nature 287,301-306.
4.Sano,H.andFeix,G.(1976)Eur.J.Biochem.71,577-583.
5.Martin,G.andKeller,W.(1998)RNA4,226-230.
Related products
Adenosine-5'-Triphosphate (ATP), 100 mM Solution77241 25 µmol
Poly(A) Polymerase, Yeast
92 888-362-2447 | 216-765-5000 | usb.affymetrix.com
Molecular Biology Enzymes | Polymerases—RNA Polymerases
Standard concentration, 20 units/µl70047Y 6,000 unitsHigh concentration, >80 units/µl70001Z 30,000 unitsCustoms by request
Source:E. colistraincontaininganoverproducingcloneofT7RNAPolymerase(1).
Description:T7RNAPolymeraseisasingle-subunitenzymeproducedbybacteriophageT7(2).ItishighlyspecificforT7promoterandterminatorsequences(2,3).Ithasbeenwidelyusedfortherapidsynthesisin vitroofspecificRNAs.ThesetranscriptscanbeuseddirectlyassubstratesforstudiesofRNAstructureorme-tabolism.Ifthetranscriptsaresuitablylabeled,theycanalsobeusedassensitivehybridizationprobes.T7RNAPolymerase,alongwithchain-terminatingnucleosidetriphosphates(4),havealsobeenusedforthedirectsequencingofDNA.T7RNAPolymeraseisalsousedtogeneratecappedmRNAforexpressionstudiesinoocytesandothercells(5).
Applications:�� ProductionofRNAtranscriptsforhybridizationprobes�� ProductionoflargeamountsofdiscretesizedRNA�� ProductionofcappedormodifiedRNAtranscripts
Properties:MolecularWeight:98.8kDaOptimumpH:7.7-8.3OptimumTemperature:37°CRequirementforDivalentCation:Mg2+(optimal
concentrationis20mM)MichaelisConstants(2):40µMATP,160µMGTP,
60µMUTP,80µMCTP
Purity:Greaterthan99%pureasdeterminedbySDS-PAGE.Testedforcontaminatingnonspecificendonucleases,exonucleases,andribonucleases.
Storage buffer:20mMHPO4,pH7.7,100mMNaCl,1mMEDTA,10mMDTT,1%CHAPS,and50%glycerol.
Assay conditions:Thereactionmixturecontains1XTranscriptionBuffer,0.5mMeachNTP,1µCilabeledCTP,1µgofT7promotercontainingDNA,andT7RNAPolymerase.Incubationisat37°Cfor15minutes(50µlreactionvolume).
Unit definition:Oneunitcatalyzestheincorporationof1nmolofCMPintopolynucleotidefractionboundtoapositivelychargedDE-81filterpaperin60minutesat37°CusingalinearizedT7promotercontaining280nttemplate.
Concentration:Standardconcentration(PN70047Y):20units/µlHighconcentration(PN70001Y/Z):>80units/µl
Functional test:TranscriptionfromplasmidcontainingT7promoter;>10µgofRNAcanbeproducedfrom1µgofsupercoiledtemplate.
Functionally tested 10X T7/T3 RNA Polym-erase Transcription Buffer (included):400mMTris-HCl,pH8.0,60mMMgCl2,100mMDTT,100mMNaCl,20mMspermidinetrihydro-chloride.
Shipping and storage:Shippedondryice.Storeat-20°C.
References:1.Tabor,S.andRichardson,C.C.(1985)Proc.Natl.
Acad.Sci.USA 82,1074-1078.2.Chamberlin,M.andRingJ.(1973)J.Biol.Chem.
248,2235-2244,2245-2250.3.Chamberlin,M.andRyan,T.(1982)inThe
Enzymes,3rdedition,ed.P.D.Boyer(AcademicPress,NewYork.) 15,87-108.
4.Axelrod,V.D.andKramer,F.R.(1985)Biochemistry 24,5716-5723.
5.Sambrook,J.andRussell,D.W.(2001)Molecular Cloning: A Laboratory Manual,3rdEdition,ColdSpringHarbor,NewYork,9.87-9.88.
6.Ausubel,F.M.,Brent,R.,Kingston,R.E.,Moore,D.D.,Seidman,J.G.,Smith,J.A.andStruhl,K.(1998)Current Protocols in Molecular Biology(JohnWileyandSons,Inc.).
T7 RNA Polymerase (RNANucleotidylTransferase,E.C.2.7.7.6)
Standard concentration, 20 units/µl70051Y 1,000 units70051Z 5,000 unitsHigh concentration, 200 units/µl70204 1,000 units 5,000 units
Source:E. coli straincontaininganoverproducingcloneofT3RNAPolymerase.
Description:T3RNAPolymerasehasahighspecificityforbacteriophageT3promotersequences.ItisusedinthesynthesisoflargeamountsofspecificRNAtranscriptsfromvectorscontainingtheT3promoter.TranscriptsmaybeusedasprobesinNorthernandSouthernblots(1),substratesforin vitro translationstudies(2),substratesforRNAprocessingstudies(3),andforexonandintronmappingofgenomicDNA(1).Theenzymemaybeusedwithbothradioactivelylabeled(4)andhapten-labeled(fluorescein,digoxigen-in,biotin,etc.)nucleosidetriphosphates(5).T3RNAPolymeraseisalsousedtogeneratecappedmRNAforexpressionstudiesinoocytesandothercells(6).
Applications:�� ProductionofRNAtranscripts�� ProductionofcappedormodifiedRNAtranscripts
Purity:Greaterthan90%pureasdeterminedbySDS-PAGE.Testedforcontaminatingnon-specificendonucle-ases,exonucleases,andribonucleases.
Storage buffer:20mMHPO4(pH7.7),100mMNaCl,1mMEDTA,10mMDTT,1.0%CHAPS,50%glycerol.
Assay conditions:Thereactionmixturecontains1XTranscriptionBuffer,0.5mMeachNTP,1µCilabeledCTP,1µgofT3promotercontainingDNA,andT3RNAPolymerase.Incubationisat37°Cfor15minutes(50µlreactionvolume).
Unit definition:Oneunitcatalyzestheincorporationof1nmolofCMPintopolynucleotidefractionboundtoapositivelychargedDE-81filterpaperin60minutesat37°CusingalinearizedT7promotercontaining280nttemplate.
Concentration:Standardconcentration(PN70051Y/Z)20units/µlHighconcentration(PN70204)200units/µl
Functionally tested 10X T7/T3 RNA Polymerase Transcription Buffer (included):400mMTris-HCl(pH8.0),60mMMgCl2,100mMDTT,100mMNaCl,20mMspermidinetrihydro-chloride.
Shipping and storage:Shippedondryice.Storeat-20°C.
References:1.Melton,D.A.,Krieg,P.A.,Regagliati,M.R.,
Maniatis,T.,Zinn,K.andGreen,M.R.(1984)Nucl.Acids Res.12,7035-7056.
2.Krieg,P.A.andMelton,D.A.(1984)Nucl.Acids Res.12,7057-7070.
3.Green,M.R.,Maniatis,T.andMelton,D.A.(1983)Cell 32,681-694.
4.Davanloo,P.,Rosenberg,A.H.,Dunn,J.J.andStudier,F.W.(1984)Proc.Natl.Acad.Sci.USA 81,2035-2039.
5.Langer,P.R.,Waldrop,A.A.andWard,D.C.(1981)Proc.Natl.Acad.Sci.USA 78, 6633-6637.
6.Sambrook,J.andRussell,D.W.(2001)Molecular Cloning: A Laboratory Manual,3rdEdition,ColdSpringHarbor,NewYork,9.87-9.88.
T3 RNA Polymerase (RNANucleotidylTransferase,E.C.2.7.7.6)
93For bulk or alternate pack sizes, email us at [email protected].
Molecular Biology Enzymes | Polymerases—Transferase
72033 500 units 2,500 unitsCustoms by request
Source:E. colicloneexpressingbovineTdT
Description:TerminalDeoxynucleotidylTransferase(TdT)catalyzesthetemplate-independentadditionofunlabelledorradio-,fluorescent-,biotin-,ordigoxygenin-labelleddeoxynucleotides,dideoxynucleotides,orribonucleo-tidestothe3'-hydroxylterminiofsingle-ordouble-strandedDNA.TdTrequiresanoligodeoxy-nucleotideprimerwithatleastthreephosphategroupsandafree3'-hydroxylendandadivalentcationforactivity.TdTalsocatalyzesthedepolymer-izationofDNA(pyrophosphorolysis)andpyrophos-phateexchange(1,2).
Historically,TdTisusedtolabelDNAprobesinapop-tosisdetectionandquantificationintheTdTdUTPnickendlabelingmethod(TUNEL)(alsoreferredtoasISEL,in situendlabeling),inDNAsequencing,(1,3)andtocreate“sticky”endsby3'tailingDNApriortoannealingandtransformation(4).For5'RACE,TdTisusedtoaddhomopolymertailstothe3'-endofthefirststrandcDNA(5).Recently,TdThasbeenusedtocloneunknownsequencesadjacenttoknownsequencesingenomicDNA(6)andforTdT-dependentPCR(7,8)(TDPCR).TDPCRisusedinRNAanaly-sis(9,10,11),transcriptionanalysis(12,13),DNAdamageanalysis(14),andDNAfootprinting(7).
Applications:�� 5'RACE�� Creatingisotopic,fluorescent,biotinylated,ordigoxygeninlabeledprobes�� Creatingfluorescent3'labeledsequencingprimers�� EliminationofbackgroundbandsinDNAsequencing�� TDPCR�� DetectionofapoptoticDNAdamage(TUNELassay)
Properties:MolecularWeight:60kDa
Purity:Testedforcontaminatingexonucleasesandendo-nucleases.
Storage buffer:50mMpotassiumphosphate,pH6.4,100mMNaCl,1mMDTT,0.1%Tween20,50%glycerol.
Assay conditions:Thereactionmixture(50µl)containing25mMsodiumcacodylate(pH7.2),0.5mMradiolabelleddATP,10mMMgCl2,0.1mMβ-ME,0.01mMd(pA)35,andenzymeisincubatedat37°Cforvaryingamountsoftime.
Unit definition:Oneunitistheamountofenzymerequiredtopolymerize1nmolofdATPresiduesintoDNAin60minutesat37°Cunderstandardassayconditions.
Concentration: 30units/µl
Functional test:Functionallytestedby3'-endlabelingofafluores-centlylabeledoligonucleotide.
Functionally tested 5X TdT Reaction Buffer (1 ml included):500mMsodiumcacodylate,pH6.8,5mMcobaltchloride,0.5mMDTT.
Shipping and storage:Shippedondryice.Storeat-20°C.
References:1. Anderson,R.S.,Bollum,F.J.,andBeattie,K.L.
(1999)Nucl. Acids Res.27,3190-3196.2. Krayevsky,A.A.,Victorova,L.S.,Arzumanov,A.
A.,andJasko,M.V.(2000)Pharm & Therapeutics85,165-173.
3. TechTip201,UseofTerminalDeoxynucleotidylTransferase(TdT)toresolveBAFLs,USBCorporation.
4. Sambrook,J.andRussell,D.W.(2001)“MolecularCloning:ALaboratoryManual,”ColdSpringHarborLaboratoryPress,ColdSpringHarbor,NY,11.110.
5. Sambrook,J.andRusselL,D.W.(2001)“MolecularCloning:ALaboratoryManual,”ColdSpringHarborLaboratoryPress,ColdSpringHarbor,NY,8.54-8.60.
6. Liu,X.andBaird,W.V.(2001)Plant Molec. Biol. Reporter19,261-267.
7. Komura,J.andRiggs,A.D.(1998) Nucl. Acids Res.26,1807-1811.
8. Davies,N.P.andMurray,V.(2000)BioTechniques29,1168-1172.
9. Schmidt,W.M.andMueller,M.W.(1996)Nucl. Acids Res.24,1789-1791.
10.Albuquerque-Silva,J.,Houard,S.,andBollen,A.(1998)Nucl. Acids Res.26,3314-3316.
11.Chen,H.,Castanotto,D.,Lebon,J.M.,Rossi,J.J.andRiggs,A.D.(2000)Nucl. Acids Res.28,1656-1664.
12.Buettner,V.L.,Lebon,J.,Gao,C.,Riggs,A.D.,andSinger-Sam,J.(2000)Nucl. Acids Res.28,e25.
13.Tagoh,H.,Himes,R.,Clarke,D.,Leenen,P.,Riggs,A.D.,Hume,D.,andBonifer,C.(2002)Genes & Development16,1721-1737.
14.Arlt,V.M.,Schmeiser,H.H.,andPfeifer,G.P.(2001)Carcinogenesis22,133-140.
Related products – Ribonucleotides
Adenosine-5'-Triphosphate (ATP), 100 mM Solution77241 25 µmol
Guanosine-5'-Triphosphate (GTP), 100 mM Solution77243 25 µmol
Related products – Deoxynucleotides
2'-Deoxyadenosine-5'-Triphosphate (dATP) Sodium Salt, 100 mM Solution77102 25 µmol
2'-Deoxycytidine-5'-Triphosphate (dCTP) Sodium Salt, 100 mM Solution77104 25 µmol
2'-Deoxyguanosine-5'-Triphosphate (dGTP) Sodium Salt, 100 mM Solution77106 25 µmol
2'-Deoxythymidine-5'-Triphosphate (dTTP) Sodium Salt, 100 mM Solution77108 25 µmol
Related products – Dideoxynucleotides
2',3'-Dideoxyadenosine-5'-Triphosphate (ddATP), 10 mM Sodium Salt Solution77110 1 µmol
2',3'-Dideoxycytidine-5'-Triphosphate (ddCTP), 10 mM Sodium Salt Solution77112 1 µmol
2',3'-Dideoxythymidine-5'-Triphosphate (ddTTP), 10 mM Sodium Salt Solution77116 1 µmol
Terminal Deoxynucleotidyl Transferase, Recombinant (rTdT)(E.C.2.7.7.31)
94 888-362-2447 | 216-765-5000 | usb.affymetrix.com
Molecular Biology Enzymes | RNase Inhibitors
71570 5,000 unitsCustoms by request
Source:Humanplacenta
Description:HumanPlacentalRNaseInhibitor(HPRI)bindsandinhibitsabroadrangeofeukaryoticRNases,includ-ingRNaseA,RNaseB,RNaseC,andhumanplacen-talRNase.HPRIispurifiedfromhumanplacentabyaffinitychromatographyonanimmobilizedRNaseAcolumn.Itformsa1:1complexwithRNaseAandinhibitsRNaseactivitynon-competitively(Ki=3x1010M).ItcanbeaddeddirectlytoreactionmixturescontainingRNA.HPRIdiffersfromothercompetitiveinhibitorsinthatitcanbeeasilyremovedfromreac-tionmixturesbyphenolextraction.
RNaseInhibitorisnoteffectiveagainstRNase1,RNaseT1,S1Nuclease,RNaseH,orRNasefromAspergillus.Tomaintainactivity,HPRIrequiresaminimumof1mMDTTandisactiveoverabroadpHrange(pH5.5-9.0).
Applications:�� cDNAsynthesis(2)
�� In vitrotranslation(3)
�� In vitrotranscription(4)
�� Polysomeisolation�� RT-PCR�� IsolationofmammaliancellsamplesthatcontainamRNA-proteincomplex(3)
�� Separationandidentificationofspecificribonucleaseactivities(5)
Properties:Activators:RequiresDTTforactivityInhibitors:RNaseswillrecoveractivityafter
inactivationattemperaturesgreaterthan55°Corin7Murea.
Purity:Testedforcontaminatingnon-specificendonu-cleases,exonucleases,ribonucleases,andlatentribonucleases.
Storage buffer:20mMHEPES-KOH(pH7.6),50mMKCl,8mMDTT,50%glycerol.
Unit definition:Oneunitinhibitstheactivityof5ngofRNaseAby50%(1).Thisinhibitoractivityisdeterminedbyitsabilitytoinhibitthehydrolysisofcyclic2'3'-CMPbyRNaseA.
Concentration:≥20units/µl
Shipping and storage:Shippedondryice.Storeat-20°C.Avoidrepeatedfreeze/thawcycles.
References:1.Blackburn,P.(1979)J. Biol. Chem.254,12484-
12487.2.deMartynoff,G.,Pays,E.andVassart,G.(1980)
Biochem. Biophys. Res. Commun.93,645-653.3.Scheele,G.andBlackburn,P.(1979)Proc. Natl.
Acad. Sci. USA76,1898-1902.4.Nielson,D.A.andShapiro,D.J.(1986)Nucl. Acids
Res.14,5936.5.Eichler,D.C.,Tatar,T.F.andLasater,L.S.(1981)
Biochem. Biophys. Res. Commun.101,396-403.
RNase Inhibitor (Human Placenta)
71571 5,000 unitsCustoms by request
Source:E. colistraincontaininganoverproducingcloneofhumanplacentaribonucleaseinhibitor.
Description:RecombinantHumanPlacentalRNaseInhibitor(rHPRI)bindsandinhibitsabroadrangeofeukary-oticRNases,includingRNaseA,RNaseB,RNaseCandhumanplacentalRNase.HPRIformsa1:1complexwithRNaseAandinhibitsRNaseactivitynon-competitively(Ki=3x1010M).ItcanbeaddeddirectlytoreactionmixturescontainingRNA.HPRIdiffersfromothercompetitiveinhibitorsinthatitcanbeeasilyremovedfromreactionmixturesbyphenolextraction.
RNaseInhibitorisnoteffectiveagainstRNase1,RNaseT1,S1Nuclease,RNaseH,orRNasefromAspergillus.Tomaintainactivity,HPRIrequiresaminimumof1mMDTTandisactiveoverabroadpHrange(pH5.5-9.0).
Applications:�� cDNAsynthesis(2)
�� In vitrotranslation(3)
�� In vitrotranscription(4)
�� Polysomeisolation�� RT-PCR�� IsolationofmammaliancellsamplesthatcontainamRNA-proteincomplex(3)
�� Separationandidentificationofspecificribonucleaseactivities(5)
Properties:MolecularWeight:50kDaIsoelectricPoint:4.7Activators:DTTInhibitors:RNaseswillrecoveractivityafter
inactivationattemperaturesgreaterthan55°Corin7Murea.
Purity:Greaterthan90%pureasdeterminedbySDS-PAGE.Testedforcontaminatingnon-specificendonucle-ases,exonucleases,andribonucleases.
Storage buffer:20mMHEPES-KOH(pH7.6),50mMKCl,8mMDTT,50%glycerol.
Unit definition:Oneunitinhibitstheactivityof5ngofRNaseAby50%(1).
Concentration:40units/µl
Shipping and storage:Shippedondryice.Storeat-20°C.Avoidrepeatedfreeze/thawcycles.
References:1.Blackburn,P.(1979)J. Biol. Chem.254,12484-
12487.2.deMartynoff,G.,Pays,E.andVassart,G.(1980)
Biochem. Biophys. Res. Commun.93,645-653.3.Scheele,G.andBlackburn,P.(1979)Proc. Natl.
Acad. Sci. USA76,1898-1902.4.Nielson,D.A.andShapiro,D.J.(1986)Nucl. Acids
Res.14,5936.5.Eichler,D.C.,Tatar,T.F.andLasater,L.S.(1981)
Biochem. Biophys. Res. Commun.101,396-403.
RNase Inhibitor (Recombinant)
95For bulk or alternate pack sizes, email us at [email protected].
Molecular Biology Enzymes | Topoisomerase
78303Y 200 units 78303Z 500 unitsCustoms by request
Source: E. coli containingacloneofthehumanTopoisomer-aseIIgene.
Description:TopoisomeraseIIaltersthetopologicalstateofnucleicacidsbypassinganintactDNAhelixthroughatransientbreakwhichgeneratesaseparateDNAhelix(1,2).Asaresultofitsdouble-strandedDNApas-sagemechanism,theenzymecanrelaxnegativelyorpositivelysupercoiledDNA,aswellascatenate/decatenateorknot/unknotDNAmolecules.Topoi-someraseIIhasanabsoluterequirementfordivalentcationandATP(ordATP).
Applications:�� RelaxnegativelyorpositivelysupercoiledDNA�� CatenatingordecatenatingDNA�� KnottingorunknottingDNA
Properties:MolecularWeight:340kDahomodimerOptimumTemperature:30°-37°COptimumpH:7.9RequirementsforDivalentCation:5mMMg2+
forcatalyticactivityand5mMCa2+forDNAcleavage.
OptimumATPConcentration:0.5to1.0mMOptimumIonStrength:100-170mMKCl/NaClInhibitors:N-ethylmaleimide,zinc,novobicin,
coumermycinA1,etoposide,amsacrine
Purity:Greaterthan98%pureasdeterminedbysilver-stainedSDS-PAGE.Freeofcontaminatingexonucle-asesandendonucleases.
Storage buffer:15mMsodiumphosphate(pH7.1),700mMNaCl,0.1mMEDTA,0.5mMDTT,50%glycerol.
Assay conditions: Thereactionmixturecontains10mMTris-HCl,pH7.9,175mMKCl,0.1mMEDTA,5mMMgCl2,2.5%glycerol,1mMATP.
Unit definition:Oneunitistheamountofenzymerequiredtofullyrelax0.3µg(5nM)ofnegativelysupercoiledpBR322plasmidDNAin15minutesat30°Cunderthestandardassayconditions.
Concentration:20units/µl
Functional test:TestedforactivityinastandardTopoisomeraseIIunitassay.
Functionally tested 10X Topoisomerse II Reaction Buffer (1 ml included):100mMTris-HCl(pH7.9),500mMNaCl,500mMKCl,50mMMgCl2,1mMEDTA,150µg/mlBSA,10mMATP.
Topoisomerase II Dilution Buffer (1 ml included):10mMsodiumphosphate(pH7.1),50mMNaCl,0.2mMDTT,0.1mMEDTA,0.5mg/mlBSA,10%glycerol.
Shipping and storage:Shippedondryice.Storeat-20°Cforfrequentuse.Storeat-80°Cforlong-termstorage.
References:1.Bjergbaek,L.,Kingma,P.,Nielsen,I.S.,Wang,Y.,
Westergaard,O.,Osheroff,N.andAnderson,A.H.(2000)J. Biol. Chem.275,13041-13048.
2.Bromberg,K.D.,Hendricks,C.,Burgin,A.B.andOsheroff,N.(2002)J. Biol. Chem.277,31201-31206.
3.Renodon-Corniere,A.,Jensen,L.H.,Nitiss,J.L.,Jensen,P.B.andSehested,M.(2002)Biochemistry41,13395-13402.
4.Sabourin,M.andOsheroff,N.(2000)Nucl. Acids. Res.28,1947-1954.
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Molecular Biology Products & KitsGene Regulation
Transfection
Nuclear Extraction
PCR Methods
Transcription Factors
Stable Cell Lines
Cellular Manipulation
NGS Library Prep
Cloning
Mutagenesis
Labeling
DNA
Markers
Protein Analysis
Proteinases
97For bulk or alternate pack sizes, email us at [email protected].
Molecular Biology Products & Kits | Table of Contents
Gene Regulation - TransfectionDeliverX™ and DeliverX Plus siRNA
Transfection Kits 98
Cellular ManipulationDeliverX Peptide Transfection Kits 99Cancer Cell Isolation Kit 101
Gene Regulation – Nuclear ExtractionNuclear Extraction Kit 102
Gene Regulation – PCR MethodsVeriQuest™ qPCR and qRT-PCR Master Mixes 103HotStart-IT™ qPCR and qRT-PCR Master Mixes 103Promoter Methylation PCR Kit 103Poly(A) Tail-Length Assay Kit 103
Gene Regulation – Transcription FactorsEMSA (Gel-Shift) Kits and Probe Sets 103TF Activation ELISA Kits 107Gene Promoter Reporter Vectors 111Luciferase Reporter Vectors 112Mammalian Full-length TF Expression Vectors 114Function-Specific Protein/DNA Arrays 115TF-TF Interaction Arrays 115Protein/DNA Arrays 116NFkB-TF and PPAR-TF Interaction Arrays 118
Gene Regulation – Stable Cell LinesReporter Stable Cell Lines 119
Next Generation Sequencing (NGS) Library PrepPrep2Seq™ DNA Library Prep Kit for
Illumina platform 123Prep2Seq Multiplex Oligo Adapters for
Illumina platform 124
CloningLigate-IT™ Rapid Ligation Kit 125
MutagenesisChange-IT™ Multiple Mutation Site Directed
Mutagenesis Kit 126
LabelingSequenase™ Random Primer Labeling Kit 127
DNA 128
DNA MarkersPCR Markers, 50-2000 bp 130DNA Ladder, 100 bp 130DNA Ladder, 1 kb Plus 131
Oligonucleotide MarkerLow Molecular Weight Marker, 10 – 100 nt 132
Protein AnalysisAngiogenesis Antibody Arrays 133Cancer Antigen ELISA Kits 134Cytokine Antibody Arrays 134
Protein MarkersProtein Markers, 10-225 kDa 136
ProteinasesProteinase K 137Proteinase K Solution 137
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Molecular Biology Products & Kits | Gene Regulation—Transfection
DeliverX siRNA Transfection Evaluation KitDX0001 0.12 mlDeliverX siRNA Transfection KitDX0002 0.4 mlDeliverX siRNA Transfection KitDX0003 1.0 mlDeliverX siRNA Transfection KitDX0004 4 x 1.0 mlDeliverX Plus siRNA Transfection Evaluation KitDX0051 0.12 mlDeliverX Plus siRNA Transfection KitDX0052 0.4 mlDeliverX Plus siRNA Transfection KitDX0053 1.0 mlDeliverX Plus siRNA Transfection KitDX0054 4 x 1.0 mlFAM-labeled siRNA ControlDX0100 0.12 mlHuman GAPDH siRNA ControlDX0101 0.06 mlDeliverX siRNA Buffer-1DX0500 4 mlDeliverX siRNA Buffer-1DX0501 14 mlDeliverX siRNA Buffer-1DX0502 34 mlDeliverX siRNA Buffer-2DX0503 4 mlDeliverX siRNA Buffer-2DX0504 14 mlDeliverX siRNA Buffer-2DX0505 34 mlDeliverX Plus siRNA Buffer-1DX0551 4 mlDeliverX Plus siRNA Buffer-1DX0552 14 mlDeliverX Plus siRNA Buffer-1DX0553 34 mlDeliverX Plus siRNA Buffer-2DX0554 4 mlDeliverX Plus siRNA Buffer-2DX0555 14 mlDeliverX Plus siRNA Buffer-2DX0556 34 ml
�� Novel peptide-based, non-endosomal delivery mechanism�� Efficient delivery of siRNA into difficult-to-transfect cell types�� High cell viability at optimal delivery conditions
DeliverX Transfection Reagents are based on novel “MPG” delivery technology developed at Centre de Recherches en Biochimie Macromoléculaire (CNRS) in Monpellier, France, in the laboratory of Dr. F. Heitz and Dr. G. Divita. MPG technology uses virus-derived amphipathic peptides that directly interact with nucleic acid cargos to form nanoparticles (150- 200 nm) capable of crossing through plasma
membranes and releasing their contents inside the cell. The mechanism of entry is receptor-independent, involves MPG/lipid interactions, and avoids the endocytic pathway, thereby preventing endosomal or lysosomal degradation of cargo. MPG peptides can be designed to accommodate specific molecular cargos including siRNAs, single and double-strand oligonucleotides, small peptides, morpholino antisense oligonucleotides, plasmids, and proteins.
DeliverX siRNA Reagents are available in two formats. DeliverX siRNA is a non-validated solution which comes with detailed instructions for carrying out assay optimization and can be used per any cell line of choice.
DeliverX Plus siRNA is a validated solution for spe-cific cell lines. With an initial offering of 18 validated cell lines, DeliverX Plus siRNA is a complete answer for your transfection needs, coming with a detailed protocol and certificate of analysis for each lot. It de-livers an assurance of success each and every time.
Regardless of which solution suits your needs, each of these reagents offers unrivalled efficiency, minimal toxicity, and a simple workflow.
Transfection proficiency controls Transfection proficiency controls are tools to enable users to optimize transfections under controlled conditions. These controls are available separately or as part of our evaluation kits.
Our FAM-labeled siRNA Control enables visualiza-tion (2–4 hours post transfection) of transfection efficiency. The human GADPH siRNA Control is a validated potent siRNA for functional evaluation of transfection efficiency.
DeliverX and DeliverX Plus siRNA Transfection Kits
U87MG cells were transfected with 10 nM of FAM-labeled siRNA Control. Two hours post transfection, bright field and fluo-rescent images of live cells were obtained.
The human brain glioblastoma astrocytoma cells were transfected with Human GADPH siRNA Control using DeliverX Plus siRNA Transfection Reagent. After 24 hours, cells were lysed and the mRNA of GADPH and cyclophilin B (internal con-trol) were quantified directly from cell lysate, without RNA purifica-tion, using QuantiGene bDNA Assay.
U87MG (human brain glioblastoma astrocytoma cells)
The differentiated 3T3-L1 mouse adipocytes were transfected with GADPH siRNA using DeliverX Plus siRNA Transfection Reagent. After 24 hours, cells were lysed and the mRNA of GADPH and cyclophilin B (internal control) were quantified directly from cell lysate, without RNA purification, using QuantiGene bDNA Assay.
Adipocytes were transfected with 10 nM of FAM-labeled siRNA Control. Two hours post transfection, bright field and fluo-rescent images of live cells were obtained.
3T3-L1 (differentiated mouse adipocytes)
Here’s how it works1. Plate cells 24 hours before
transfection.
2. Form transfection complex Prepare siRNA/Buffer-1. Prepare Transfection Reagent/
Buffer-2. Combine siRNA/Buffer-1 and
Transfection Reagent/Buffer-2. Incubate complex at 37°C for
20 minutes.
3. Transfect cells/quantity knockdown
Add complex, incubate 3–5 minutes at RT.
Add serum-free media, incubate in 5% CO2 for 2–4 hours.
Add complete medium, incubate in 5% CO2 for 24–72 hours.
Quantify mRNA and/or protein knockdown.
99For bulk or alternate pack sizes, email us at [email protected].
Molecular Biology Products & Kits | Gene Regulation—Transfection/ Cellular ManipulationDeliverX and DeliverX Plus siRNA Transfection Kits continued...
The human THP-1 suspension cells were transfected with Human GADPH siRNA Control using DeliverX Plus siRNA Transfection Reagent. After 24 hours, cells were lysed and the mRNA of GADPH and cyclophilin B (internal control) were quantified directly from cell lysate, without RNA purification, using QuantiGene bDNA Assay.
THP-1 (human peripheral blood acute mono-cytic leukemia cells)
HeLa cells were transfected with Human GADPH siRNA Control using DeliverX siRNA Transfection Reagent. After 24 hours, cells were lysed and the mRNA of GADPH and cyclophilin B (internal control) were quantified directly from cell lysate, without RNA purification, using QuantiGene bDNA Assay.
HeLa (human cervical adenocarcinoma)
The differentiated C2C12 mouse myotubes were transfect-ed with GADPH siRNA using DeliverX Plus siRNA Transfection Reagent. After 24 hours, cells were lysed and the mRNA of GADPH and cyclophilin B (internal control) were quantified directly from cell lysate, without RNA purification, using QuantiGene® bDNA Assay.
Myotubes were transfected with 10 nM of FAM-labeled siRNA Control. Two hours post transfection, bright field and fluo-rescent images of live cells were obtained.
C2C12 (differentiated mouse myotubes)
DeliverX TAMRA-labeled Peptide Control DX1100 75 µlDeliverX Pre-formed Peptide Transfection Complex ControlDX1101 1.5 mlDeliverX Peptide Transfection Evaluation KitDX1001 0.6 ml*DeliverX Peptide Transfection KitDX1002 3.0 mlDeliverX Peptide Buffer-1DX1500 4 mlDeliverX Peptide Buffer-1DX1501 20 mlDeliverX Peptide Buffer-2DX1502 4 mlDeliverX Peptide Buffer-2DX1503 20 ml
* DeliverX Peptide Evaluation Kits are configured for new users and contain the following:
a. 0.6 ml DeliverX Peptide Transfection Reagentb. 75 µl DeliverX TAMRA-labeled Peptide Control,c. 1.5 ml DeliverX Pre-formed Peptide Transfection
Complex Control
�� Novel peptide-based, non-endosomal delivery mechanism�� Efficient delivery of a wide variety of peptides into most cell types�� Versatile—form a single transfection complex and dilute to evaluate multiple peptide concentrations�� High cell viability at optimal delivery conditions
Delivery of peptides into cells has traditionally been difficult to accomplish, both in terms of efficiency and retaining normal cellular function post transfec-tion. With the introduction of the DeliverX Peptide Transfection Kit, it is now possible to reproduce and easily deliver a wide variety of peptides into many cell types. Peptide transfection is being pursued for various applications, including modulation of cell signaling pathways for basic research, as potential therapeutic agents, and for the study of the conse-quences of pseudo up-regulation of specific cellular peptides.
Here’s how it worksDeliverX Transfection Reagents are based on novel ”MPG“ delivery technology developed at Centre de Recherches en Biochimie Macromoléculaire (CNRS) in Montpellier (France) in the laboratory of Dr. F. Heitz and Dr. G. Divita.
MPG technology uses virus-derived amphipathic peptides that directly interact with nucleic acid cargos to form nanoparticles (150-200 nm) capable of diffusing through plasma membranes and releas-ing their contents inside the cell. The mechanism of entry is receptor-independent, involves MPG/lipid membrane interactions, and avoids the endocytic pathway, thereby preventing endosomal or lysosomal degradation of cargos.
DeliverX Peptide Transfection Kits
1. MPG peptide interacts with peptide to form nanoparticle.
2. MPG/peptide nanoparticle interacts with the external side of the cellular membrane.
3. The MPG/peptide particle inserts into the membrane (a pore-like structure is created).
4. MPG/peptide particle is internalized and the peptide is decoupled from the complex.
(Continued on next page)
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Molecular Biology Products & Kits | Cellular Manipulation
Transfection of FITC-labeled Cdk2 Peptide Inhibitor
Delivery of FITC-labeled Cdk2 peptide inhibitor. FITC-labeled Cdk2 peptide inhibitor was transfected into HeLa cells using DeliverX Peptide Transfection Reagent.
MPG peptides can be designed to accommodate specific molecular cargos including siRNAs, single and double-strand oligonucleotides, small peptides, morpholino antisense oligonucleotides, and plasmids.
Inhibition of cell proliferationCyclin dependent kinase (Cdk) 2 is required at mul-tiple stages for progression through the mammalian cell cycle. In association with cyclin E, Cdk2 pro-motes the transition from G1 to S phase, and Cdk2/cyclinA complexes drive cells through S phase. The activities of Cdks are highly regulated by association with cyclins, whose temporal expression throughout the cell cycle is tightly controlled by phosphorylation or dephosphorylation by kinases and phosphatases, and through the actions of inhibitory proteins such as Ink4 and Cip/Kip. These key regulators of mammalian cell proliferation are excellent targets for anticancer agents. Here we demosnstrate that cellular proliferation can be inhibited by transfection of a Cdk2 inhibitory peptide into HeLa cells using the DeliverX Peptide Transfection Reagent.
Easy-to-use kitsDeliverX Peptide Transfection KitsOur DeliverX Peptide Transfection Kits are suitable for transfection of most cell types. DeliverX Peptide Transfection Kits contain the reagents required to efficiently transfect peptides into most cell types with minimal cell damage and good reproducibility when following the optimization guidelines provided in the user manual.
Transfection proficiency controlsTransfection proficiency controls are tools to enable users to optimize transfections under controlled conditions. These controls are available separately or as part of our evaluation kit.
DeliverX TAMRA-labeled Peptide Control enables visualization of transfection efficiency 1–4 hours post transfection. This validated peptide control is a 15 amino acid peptide with the TAMRA fluorophore at the carboxyl terminus. The quantity provided is sufficient for ½ of one 96-well plate.
DeliverX Pre-formed Peptide Transfection Complex Control—Enables you to test whether the DeliverX Peptide Transfection Reagent is compat-ible with your cell line or cell type. The Peptide Trans-fection Complex Control consists of the pre-formed complex of DeliverX Peptide Transfection Reagent and the TAMRA-labeled Peptide Control. The quan-tity provided is sufficient for ½ of a 96-well plate.
DeliverX Peptide Transfection Kits continued...
Cell only, time = 48 hours
DeliverX Peptide Transfection Reagent + Nega-tive Control Peptide, time = 48 hours
DeliverX Peptide Transfection Reagent + Cdk2 Peptide Inhibitor, time = 48 hours
Delivery of a Cdk2 peptide inhibitor to inhibit cell-cycle progression. HeLa cells grown in a 96-well microplate were transfected with a Cdk2 peptide inhibitor using DeliverX Peptide Transfection Reagent and incubated for 48 hours.
101For bulk or alternate pack sizes, email us at [email protected].
Molecular Biology Products & Kits | Cellular Manipulation
Cancer Cell Isolation Kit (2)CI0002 2 assaysCancer Cell Isolation Kit (4)CI0004 4 assaysCancer Cell Isolation Kit (10)CI0010 10 assays
�� Purify cancer cells from fresh biopsy samples�� Establish primary cell lines or use for biochemical assays�� Ideal for proteomics applications
The Cancer Cell Isolation Kit provides a simple, cost-effective method for isolating cancer cells from biopsy samples. Cancer cells purified using this technique can be used immediately for a wide range of applications, including biomarker identification, microarray hybridization, and genetic analysis. More-over, the whole cell isolation procedure is so gentle that cells can be used to establish primary cell lines.
Purify any population of cancer cells The ability to obtain uncontaminated cancer cell samples is one of the major bottlenecks in the study of tumor development and cancer biology. Tumor biopsies from cancer patients and animal tumor models often contain a heterogeneous population of cells that include normal tissue, blood, and cancer cells. This mixed population makes diagnosis and valid experimental conclusions difficult to obtain and interpret.
Typically, cancer cells are isolated from biopsy samples via microdissection, a time-consuming technique that requires costly equipment and surgical precision. Another conventional method is immunoprecipitation with antibodies specific to each cancer cell type. Unlike these techniques, the Cancer Cell Isolation Kit requires no specialized skill set and no expensive equipment, and can be used to isolate many cancer cell types.
Simple, cost-effective techniqueThe cancer cell isolation method only takes a couple of hours, and involves just a few rounds of incuba-tion, washing, and centrifugation. Using the same amount of starting biopsy material required for conventional methods, this technique can be used to isolate a much greater quantity of cancer cells.
Cancer cells purified using the Cancer Cell Isolation Kit can be used directly for proteomics applications such as identification of biomarkers or for RNA-based applications such as microarray hybridization and genetic analysis. Cells can also be used to purify proteins to test antibody-based cancer therapies. Alternatively, cells can be used to establish primary cell lines.
To prepare nuclear extracts from isolated cancer cells, please see the Nuclear Extraction Kit, PN AY2002 on page 102.
Kit components:Tumor cell suspension solutionTumor cell digestion solutionTumor cell purification solution100 µm filter
Cancer Cell Isolation Kit
Here’s how it works
(Continued on next page)
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Molecular Biology Products & Kits | Cellular Manipulation/Nuclear Extraction
Cancer Cell Isolation Kit continued...
AY2002 * *20 rxn from cell culture; 10 rxn from whole tissue
The Nuclear Extraction Kit is ideal for preparing nuclear extracts from cell culture or tissues for use with Protein/DNA Arrays or electrophoretic mobility shift assays (EMSA). This kit facilitates the extraction of functional crude nuclear proteins from various cell types or whole tissue.
Here’s how it worksThe protocol is optimized for approximately 107 cells (near-confluent 100 mm plate) or 0.5 g of tissue. First, cells are allowed to swell with hypotonic buffer. Then, the cells are disrupted, nuclei are collected, and nuclear proteins released with a high-salt buffer.
Kit components:Buffer ABuffer B100 mM DTTProtease inhibitorPhosphatase inhibitor IPhosphatase inhibitor II
Nuclear Extraction Kit
103For bulk or alternate pack sizes, email us at [email protected].
EMSA KitAY1XXX kitEMSA TF Probe SetsAY1XXXP* eaEMSA Combo Kit (any three probes)AY9999 kitEMSA Kit without Probe Set AY1000 kit*Use 4 digits from EMSA Prod. No.See chart on following page for full listing of EMSA kits.
EMSA (Electrophoretic Mobility Shift Assay) Kits* are useful tools for validating results from Protein/DNA Arrays to identify proteins that interact with DNA. You can also use them to assess the DNA-binding activity of a specific transcription factor. This rapid technique is based on the different electrophoretic mobilities of free DNA and protein/DNA complexes in native (non-denaturing) polyacrylamide gels.
Here’s how it works1. Incubate the biotin-labeled DNA probe with
your nuclear extracts which should contain your protein(s) of interest.
2. Separate the mixture on a nondenaturing polyacrylamide gel.
3. Transfer the protein/DNA complex to a nylon membrane. Incubate the blot with streptavidin-HRP and then add chemiluminescent substrate.
4. Visualize shifted bands using film or chemiluminescence imaging system that correspond to the protein DNA complexes.
To confirm whether the DNA-binding activity of a particular protein is specific, you simply add an excess of cold, unlabeled DNA probe to the reaction mixture. If binding is specific, the cold probe com-petes with the labeled DNA probe for binding to the protein. As a result, the shifted band is eliminated.
EMSA probe setsIn addition to our EMSA Kits, we offer individual EMSA probes for purchase without the other EMSA Kit components. EMSA TF Probe Sets are intended for use with our EMSA Kits. Each TF probe set is pro-vided as 25 µl (10 ng/µl) of biotin-labeled TF probe accompanied by 25 µl (330 ng/µl) of cold probe, its unlabeled form.
EMSA nomenclature For some Transcription Factors, multiple consensus sequences for the same TF have been published in the literature and, where appropriate, we have provided those variations available in our EMSA kits and arrays. For the TF’s with multiple consensus sequences, they have been denoted with numbers in parenthesis, i.e. (1), (2), (3) etc. Also, PR = Promoter and BP = Binding Protein. To obtain the consensus sequences and references for the TFs used in our arrays, please contact our Technical Support group.
Kit components:Biotin-labeled and cold TF probesBiotin-labeled and cold control probesControl nuclear extractBinding and detection reagentsEach EMSA Kit is supplied with enough reagents to run 25 reactions.
See chart on the following page for a full listing of EMSA kits.
To prepare nuclear extracts for this procedure, please see the Nuclear Extraction Kit, PN AY2002 on page 102.
EMSA (Gel-Shift) Kits and Probe Sets
PCR Methods
Molecular Biology Products & Kits | Gene Regulation— PCR Methods/Transcription Factors
(Continued on next page)
Affymetrix offers a portfolio of USB branded products using PCR methods to study or quantify gene regulation events. Full descriptions of these products can be found in the PCR Tools or RNA Analysis sections of the catalog. Below is a brief summary:
Product Description
VeriQuest qPCR and qRT-PCR Master Mixes Our newest family of hot start qPCR master mixes, the VeriQuest product line, uses our own chemically modified Taq polymerase in specially formulated ready-to-use master mixes. If you’re looking for PCR efficiency and robustness which exceed the current gold standards, these mixes are for you. See pages 26-44.
HotStart-IT qPCR and qRT-PCR Master Mixes Our Hotstart-IT master mixes use a novel, patented primer sequestration method for hot start qPCR and qRT-PCR. If you’re looking for a qPCR mix with faster, easier enzyme activation, give this product a try. See pages 26-44.
Promoter Methylation PCR Kit Epigenetic analysis of your favorite promoter(s) made easy with a non-bisulfite conversion methodology. See page 24.
Poly(A) Tail-Length Assay Kit Did you know that polyadenylation length impacts the fate and function of an mRNA? Here’s an innovative kit to measure poly(A) tail-length which is far easier than the Northern blot approach. See page 158.
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Molecular Biology Products & Kits | Gene Regulation—Transcription Factors
EMSA (Gel-Shift) Kits and Probe Sets continued...
AAF AY1259
ABF-1 AY1260
ACF AY1261
ACP BP AY1262
ADD-1 AY1158
ADR-1 AY1059
AF-1 AY1159
AFP-1 AY1160
Afxh/Foxo-4 AY1087
AhR/Arnt AY1060
AIC/CBF AY1161
AIC-2/3/4 AY1263
ALF-1 AY1264
α PAL AY1265
AML-1 AY1162
ANG/IRE AY1061
Antioxidant RE AY1062
AP-1 (1) AY1001
AP-1 (2) AY1055
AP-2 (1) AY1002
AP-2 (2) AY1056
AP-2 (3) AY1266
AP-2/YY1 AY1163
AP-3 (1) AY1063
AP-3 (2) AY1267
AP-4 (1) AY1064
AP-4 (2) AY1268
apo A1 PR AY1269
AR AY1003
AREB-6 AY1164
ARP AY1165
ATF (1) AY1270
ATF (2) AY1167
ATF (3) AY1271
ATF/CRE AY1166
beta-M-globin
factor B1 AY1272
beta-RE AY1065
Brn-3 AY1004
BZP AY1273
CACC AY1168
CBF (1) AY1006
CBF (2) AY1279
CBFB AY1278
CCAAT AY1169
CCAC AY1170
CD28RC (1) AY1171
CD28RC (2) AY1172
CDP AY1007
CdxA/NKX2 AY1156
CEA AY1174
C/EBP AY1005
C/EBPαγ AY1274
CEBPα(1) AY1275
CEBPα(2) AY1276
C/EBPγ AY1277
CEF-1 AY1066
CEF-2 AY1067
CETP/CRE AY1068
c-Ets-1 AY1280
c-fos BP AY1281
c-Myb (1) AY1008
c-Myb (2) AY1282
c-Myc AY1283
COUP-TF (1) AY1069
COUP-TF (2) AY1284
CP-1 AY1286
CP1/CTF/CBTF AY1175
CP-1B AY1285
CP-2 AY1287
CPE AY1176
CRE AY1289
CREB (1) AY1009
CREB (2) AY1288
CREB-2 AY1177
CREB-BP-1 AY1070
c-Rel AY1071
CSBP AY1178
CTCF AY1179
CYP1A1 AY1290
DE-1 AY1291
Dioxin Receptor AY1292
DR-0 AY1073
DR-1 AY1074
DR-2 AY1075
DR-3 AY1076
DR-4 AY1077
DR-5 AY1078
E12 (1) AY1180
E12 (2) AY1295
E12 (3) AY1293
E12/E47 AY1294
E2 AY1297
E2F-1 (1) AY1010
E2F-1 (2) AY1296
E-47 AY1079
E4BP-4 (1) AY1080
E4BP-4 (2) AY1298
E4F/ ATF AY1081
EBP-40/45 AY1181
EBP-80 AY1299
EBP-C2/NF-
muE3/YEB3 AY1300
EGF BP AY1301
EGR-1 (1) AY1011
EGR-1 (2) AY1303
EGR-1 (3) AY1302
EGR-2 (1) AY1304
EGR-2 (2) AY1305
EIL-1/2/3 AY1306
EKLF (1) AY1182
EKLF (2) AY1183
ELF AY1184
Elk-1 AY1082
ER AY1012
ETF AY1307
Ets AY1013
Ets-1/PEA-3 AY1014
ETV4/E1A-F AY1308
EVI-1 AY1083
FAST-1 AY1015
FKHR (1) AY1085
FKHR (2) AY1086
Freac-2 (1) AY1088
Freac-2 (2) AY1084
Freac-4 AY1089
Freac-7 AY1090
GAG AY1091
GAS AY1092
GAS/ISRE AY1016
Gastrin EGF RE AY1310
GATA AY1017
GATA-1 (1) AY1093
GATA-1 (2) AY1311
GATA-1/2 AY1094
GATA-2 AY1095
GATA-3 AY1096
GATA-4 AY1097
GATA-6 AY1098
GBF-1/2/3/HY5 AY1312
GCF AY1313
GFI-1 AY1099
GKLF AY1314
GR/PR AY1018
H1TF-2 AY1315
H4TF AY1185
H4TF-1 AY1316
HAS AY1108
HBS (1) AY1100
HBS (2) AY1157
HEN-1 AY1186
HFH-1 (1) AY1102
HFH-1 (2) AY1318
HFH-11β/11α AY1317
HFH-2 AY1103
HFH-3 AY1104
HFH-8 AY1105
HFH-8/3/LUN AY1187
HFS/HAS AY1101
HIF-1 AY1107
HiNF AY1188
HiNF/D3 AY1322
HiNF-A AY1319
HiNF-B/H1TF1 AY1320
HiNF-C AY1321
HLF AY1189
HMG AY1190
HNF-1 (1) AY1191
HNF-1 (2) AY1324
HNF-1 a/b/c AY1323
HNF-1α AY1192
HNF-3 (1) AY1109
HNF-3 (2) AY1110
HNF-4 (1) AY1019
HNF-4 (2) AY1111
HNF-4/COUP-TF AY1112
HNF-4α2/1 AY1325
HOX4C AY1193
HOXA-4 AY1326
HOXD-8 (1) AY1195
HOXD-8 (2) AY1196
HOXD-8/9/10 AY1194
HOXD-9/10 AY1197
HSE AY1020
HSF-2 distal AY1327
HSF-3 proximal AY1328
ICSBP AY1198
IFI-56KBP AY1330
Ikaros AY1114
IL6-RE-BP AY1331
IR-4/EBV BP AY1332
IRF-1 AY1021
IRF-1/2 AY1115
ISGF AY1333
Isl-1 AY1199
ISRE (1) AY1116
ISRE (2) AY1117
ISRE (3) AY1118
kBF-α AY1334
KKLF AY1119
KPF-1 AY1200
KTP-1 AY1201
lactoferrin BP AY1335
LCR-F1 AY1202
LEF-1 AY1203
LF-A1 (1) AY1204
LF-A1 (2) AY1336
LF-A2 AY1337
LF-B2 AY1338
LH2/Lim1 AY1339
L-III BP AY1120
LR-1 AY1205
LSF AY1340
LXRE-1 AY1341
LyF-1 (1) AY1206
LyF-1 (2) AY1207
LyF-1 (3) AY1208
MASH-1 AY1342
MAZ AY1209
MAZ (1) AY1210
MBP-1 (1) AY1343
MBP-1 (2) AY1344
mck PR E1 AY1345
mck PR E2 AY1346
MDBP (1) AY1S47
MDBP (2) AY1348
MEF-1 AY1022
MEF-2 AY1023
MEF-2(1) AY1121
MEF-2(2) AY1122
MEF-2a AYI349
MEF-3 AY1123
Mfh-1 AY1350
MHC gene PR
W Box AY1351
MRE AY1024
MSP-1 AY1124
msx-1/2/3 AY1352
hTERT-MT-Box AY1113
MTB-Zf AY1211
MTF (1) AY1212
MUSF-1 AY1125
Myb (2) AY1353
myc/CF1 AY1354
Myc-Max AY1025
MyoD AY1213
MyoG AY1214
MyTI AY1215
MZF-1 AY1126
MZF-1(1) AY1216
MZF-1(2) AY1217
MZF-1(3) AY1218
NCAM-BP AY1356
NF-1 AY1026
NF-1 (2) AY1357
NF-1/L AY1358
NF-4FA AY1219
NF-A AY1360
NF-A3 AY1359
NFAT-1 (1) AY1027
NFAT-1 (2) AY1361
NF-Atp AY1362
NF-Atx AY1363
NF-E1/YY1 AY1028
NF-E2 (1) AY1029
NF-E2 (2) AY1364
NF-E6/CP1 AY1365
NF-Gma (1) AY1220
NF-Gma (2) AY1221
NF-IL-2 AY1222
NFκB (1) AY1030
NFκB (2) AY1366
NFκB (3) AY1367
NF-Y (1) AY1127
NF-Y (2) AYI223
Nkx-2.5 AY1128
NPAS-2 AY1129
N-ras BP AY1368
NRF-1 AY1224
NRF-2 AY1225
NZF-3 AY1131
OBP AY1369
OCT (1) AY1031
OCT (2) AYI370
OCT (3) AY1371
OCT (4) AY1372
ODC AY1373
ORE AY1132
p300 AY1133
p53 (1) AY1032
p53 (2) AY1374
p53 (3) AY1226
p55 AY1227
PARP AY1134
PAX-1 AY1375
PAX-2 (1) AY1135
PAX-2 (2) AY1376
PAX-3 AY1136
PAX-4 AY1137
PAX-5 (1) AY1033
PAX-5 (2) AY1377
PAX-6 (1) AY1138
PAX-6 (2) AY1378
PAX-8 AY1139
PBGD BP AY1379
Pbx-1 AY1034
PEA3/HIP AY1228
PEBP AYI229
PEBP-2 (1) AY1230
PEBP-2 (2) AY1381
PEPCK PR GR AY1382
Pit-1(1) AY1035
Pit-1(2) AY1383
Pit-1(3) AY1384
PO-B AY1385
PPAR (1) AY1036
PPAR (2) AY1140
PPAR (3) AY1141
PPUR (1) AY1231
PPUR (2) AY1232
PRDI-BFc AY1386
PRDII-BF1 AY1387
PRE AY1037
PREB AY1388
PTF-1 AY1390
PTF-1β AY1389
PU.1 AY1391
PuF AY1233
PUR AY1392
Pur-1 AY1234
PYR AY1235
RAR/DR-5 AY1038
RAR/RXR/CF1/
MB67 AY1393
RB AY1236
RFX-1/2/3 (1) AY1237
RFX-1/2/3 (2) AY1238
RFX-1/2/3 (3) AY1394
RIPE-3a1 AY1239
RORE AY1395
RREB (1) AY1142
RREB (2) AY1143
RSRFC4 AY1144
RVF AY1396
RXR/DR-1 AY1039
SAA AY1145
SAP-1b AY1397
SIE AY1040
SIF-1 AY1240
SIF-2 AY1241
SIF-3 AY1242
Skn AY1146
SMAD/SBE AY1041
SMAD-3/4 AY1042
Snail AY1398
Sp-1 (1) AY1043
SP-1 (2) AY1243
Sp-1 (3) AY1399
SPERM-1 AY1400
SRE AY1044
SRF (1) AY1402
SRF (2) AY1401
SRF/SAP AY1244
SRY AY1147
SRY (2) AY1245
SSAP AY1403
STAT-1 AY1045
STAT-3 AY1046
STAT-4 AY1047
STAT-5 AY1048
STAT-5b AY1405
STAT-6 AY1049
Surf-2 (1) AY1406
Surf-2 (2) AY1407
T3R AY1408
Tat AY1409
Tax/CREB AY1148
TCE AY1246
TCF/LEF AY1149
TEF-1 AY1411
TEF-1/AP-5 AY1410
TFE-3 AY1247
TFE-3L AY1412
TFEB AY1413
TFIID AY1050
Tf-LF AY1248
TGT-3 AY1249
Thy-1 BP AY1414
TIF-1 AY1250
tPA BP AY1415
TR AY1051
TR/DR-4 AY1052
Transferrin BP AY1416
TREF-1/2 AY1251
TTF-1 (1) AY1417
TTF-1 (2) AY1418
TxREF/NF-lll AY1419
URE AY1420
USF-1 (1) AY1053
USF-1 (2) AY1421
VDR/DR-3 AY1054
v-Maf AY1252
v-rel 50-55K AY1422
WAP BP AY1423
WT1 (1) AY1253
WT1 (2) AY1254
WT1 (3) AY1255
X2 BP AY1256
XBP-1 (1) AY1150
XBP-1 (2) AYI424
XBP-1/X2 BP AY1257
XRF AY1151
XRE AY1425
YB-1 AY1426
Yi AY1427
ZEB AY1428
ZIA AY1152
ZIB AY1153
ZIC AY1154
ZID AY1155
Zn15 AY1429
ZNF174 AY1258
TF Prod. no. TF Prod. no. TF Prod. no. TF Prod. no. TF Prod. no. TF Prod. no. TF Prod. no.
Fra-1/JUN AY1309 HSF/HTF AY1329 myc-PRF AY1355 PCF AY1380 STAT-1/3 AY1404
105For bulk or alternate pack sizes, email us at [email protected].
1 2 3
1 2 3
AP-1 The activator protein 1 (AP-1) transcription factor is composed of heterodimers of the Fos-activating transcription factor (ATF) and Jun subfamilies of proteins. The composition and activity of these com-plexes is regulated by the MAPK signaling pathway. AP-1 binds to transcription response elements in the promoter region of many genes involved in cell proliferation.
Lane 1. Probe Lane 2. Probe + PMA treat. HeLa nuc. ext. Lane 3. Probe + PMA treat. HeLa nuc. ext. + cold probe.
EREstrogen Receptor a (ERa) is activated by binding to estrogen that causes a conformational change and releases ERa from its inhibitory complex. ERa then binds to the estrogen response element of tar-get genes, activating gene transcription. This struc-tural change is also induced by phosphorylation of ERa, enabling substantial “cross-talk” between growth factor and estrogen receptor signaling.
Lane 1. Probe Lane 2. Probe + PMA treat. HeLa nuc. ext. Lane 3. Probe + PMA treat. HeLa nuc. ext. + cold probe.
EtsThe Ets-1 proto-oncogene is a member of the Ets family that demonstrates homology to the v-ets oncogene. Ets activity is regulated both by phosphorylation and via the formation of various transcription factor complexes. Activated Ets-1 binds to the Ets-binding motif and promotes expression of genes that control cell proliferation, angiogen-esis, apoptosis, and differentiation.
Lane 1. Probe Lane 2. Probe +HeLa nuc. ext.Lane 3. Probe + HeLa nuc. ext. + cold probe.
CREBThe transcription factor cAMP response element binding protein (CREB) is activated by phos-phorylation in response to a variety of extracel-lular signals, including growth factors, hormones, and neurotransmitters. Active CREB binds to the cAMP-responsive element, causing transcription of multiple genes.
Lane 1. Probe Lane 2. Probe + PMA treat. HeLa nuc. ext. Lane 3. Probe + PMA treat. HeLa nuc. ext. + cold probe.
EGR-1Early growth-response factor 1-(EGR-1) expression/activation is induced by signals that stimulate mito-genesis and differentiation, as well as by changes in the local cellular environment. EGR-1 interacts with a G+C-rich consensus-binding site and induces expression of many genes important in inflamma-tion, cell growth, apoptosis, and the pathogenesis of disease.
Lane 1. ProbeLane 2. Probe + Jurkat nuc. ext.Lane 3. Probe + Jurkat nuc. ext. + cold probe.
GR Glucocorticoid receptor (GR) is a nuclear hormone receptor. Upon glucocorticoid binding to GR, a change in receptor conformation occurs, causing it to dissociate from regulatory heat shock proteins. GR then translocates to the nucleus, where it binds to glucocorticoid response elements (GREs) to modulate expression of genes involved in metabolic, cardiovascular, immune, and behavioral functions.
Lane 1. ProbeLane 2. Probe + Dex. treat. HeLa nuc. ext.Lane 3. Probe + Dex. treat. HeLa nuc. ext. + cold probe.
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Molecular Biology Products & Kits | Gene Regulation—Transcription Factors
Our 12 Most Popular EMSA Kits
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PPAR (1)Peroxisome proliferator activated receptors (PPARs) are regulated by small molecules such as plasticiz-ers, herbicides, unsaturated lipids, eicosanoids, fibrates, and thiazolidinediones. Phosphorylation of PPAR by MAP kinase causes increased transcrip-tional activity, whereas phosphorylation of PPAR by MAP kinases inhibits transcriptional activity.
Lane 1. ProbeLane 2. Probe + PMA treat. HeLa nuc. ext.Lane 3. Probe + PMA treat. HeLa nuc. ext. + cold probe
NFkB (1)The activity of NFkB is tightly regulated by interac-tions with IkB proteins. A variety of stimuli such as cytokines, growth factors, viruses, and environmen-tal hazards can trigger activation of IkB kinases, which phosphorylate IkB. This induces a conforma-tional change in IkB; NFkB then dissociates from the complex, translocates into the nucleus, and mediates transcriptional activation of target genes.
Lane 1. ProbeLane 2. Probe + TNFa treat. HeLa nuc. ext.Lane 3. Probe + TNFa treat. HeLa nuc. ext. + cold probe
p53The tumor suppressor p53 is maintained at low lev-els in cells. DNA damage induces phosphorylation or acetylation of p53, causing its stabilization and accumulation in the nucleus. p53 has the potential to act as a transcriptional activator or repressor, inducing expression of downstream genes that inhibit the cell cycle or induce apoptosis.
Lane 1. Probe.Lane 2. Probe + A431 nuc. ext.Lane 3. Probe + A431 nuc. ext. + cold probe
STAT-1The STAT-1 transcription factor undergoes tyrosine phosphorylation in response to IFNγ binding to its cognate receptor. This phosphorylation causes STAT-1 to dimerize and translocate to the nucleus, where it binds to the gamma interferon-activated site (GAS) promoter and induces the transcription of IFNγ-dependent genes. STAT-1 can also be phos-phorylated by numerous serine/threonine kinases, which increase transcriptional activity.
Lane 1. ProbeLane 2. Probe + A431 nuc. ext.Lane 3. Probe + A431 nuc. ext. + cold probe
HIF-1Hypoxia-inducible factors (HIFs) are activated by low-oxygen conditions. HIF proteins are extremely unstable in the presence of oxygen due to a high degradation rate by the proteasome. Hypoxia dramatically increases the half-life of HIF, which induces regulation of most genes activated by hypoxia.
Lane 1. ProbeLane 2. Probe + PMA treat. HeLa nuc. ext.Lane 3. Probe + PMA treat. HeLa nuc. ext. + cold probe
NFATNuclear factor of activated T cells (NFAT) is activated when T-cell receptors are stimulated. T-cell activation in turn activates calcineurin, the phosphatase that dephosphorylates NFA. This induces a conformational change in NFAT, which causes it to translocate to the nucleus, where it binds to NFAT response elements. NFAT target genes are involved in the regulation of immune responses, differentiation, and apoptosis.
Lane 1. ProbeLane 2. Probe + CD3 treat Jurkat nuc. ext.Lane 3. Probe +CD3 treat. Jurkat nuc. ext.+ cold probe
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Our 12 Most Popular EMSA Kits continued...
Molecular Biology Products & Kits | Gene Regulation—Transcription Factors
107For bulk or alternate pack sizes, email us at [email protected].
See chart on page 110 for a full listing of TF Activation ELISA Kits
�� Quantify activation of AP-1, E2F-1, EGR1, ERa, FKHR, GATA-1, GR, HIF-1a, HNF3b, MEF-2, MITF, NFkB c-Rel, NFkB p50, NFkB p52, NFkB p65, NkKB Rel-b, p53, PPARa, PPARγ, SMAD-2, SMAD-4, SP-1, and STAT-1�� Detect activation with as low as 0.5 µg nuclear extract �� Antibody and double-stranded DNA cis element give assay the best specificity and sensitivity possible�� Simple workflow: entire assay takes less than 4 hours
The TF Activation ELISA Kits are useful tools to detect and quantify transcription factor activation. The ELISA-based kits combine a fast, user-friendly format with a sensitive and specific assay. The assay detects activation with as low as 0.5 µg of cellular extract and the whole assay is completed in fewer than 4 hours.
An EMSA or gel shift assay typically requires 5 µg of nuclear extract and does not have the level of specificity of a TF ELISA which uses an antibody directed to the TF of interest. Each 96-well plate is comprised of individual strips of 8 wells suitable for low or high-throughput applications. These kits may be used to study drug potency, inhibitor or activator proteins, or to probe protein structure and function in signaling pathways.
Kit components:TF Antibody and HRP-conjugated secondary
antibodyConsensus DNATMB colorimetric reagent 96-well assay platePositive control nuclear extract or recombinant
protein
Here’s how it works:The TF ELISA kits use a proprietary, patent-pending technology developed for quantification of transcrip-tion factor activation. The entire procedure can be completed in less than 4 hours.
Three basic steps are involved:1. An oligonucleotide containing an TF consensus
binding site is immobilized on the 96-well plate.2. Activated TF from nuclear or whole-cell extracts
specifically binds to this oligonucleotide.3. The complex bound to the oligonucleotide is
detected by an antibody directed against the TF subunit. An additional secondary HRP-conjugated antibody provides sensitive colorimetric readout easily quantified by spectrophotometry.
TF Activation ELISA Kits
TF ELISA Kits are ideal for quantifying transcription factor binding to DNA.
PPARa ELISA KitThe PPARa ELISA Kit quantifies PPARa transcription factor activation.
ERa ELISA KitThe ERa ELISA Kit quantifies ER transcription factor activation.
Typical results obtained by using the PPARa ELISA kit. Different amounts of nuclear extract as indicated were used for the assay. Cos-1: nuclear extract from Cos-1 cells incubated overnight in DMEM + 0.1% FBS, untreated. Cos-1+PPARa+RXRA: nuclear extract from Cos-1 cells (6–8 x 106 cells/dish) co-transfected over-night (16 hours) with 5 µg CMV-PPARa and 5 µg CMV-RXRA expres-sion vectors.
NFkB p50 ELISA KitThe NFkB p50 ELISA Kit quantifies NFkB p50 transcription factor activation.
Typical results obtained by using the ERa ELISA kit. Different amounts of nuclear extract as indicated were used for the assay. MCF7: nuclear extract from MCF-7 cells grown in DMEM + 10% FBS. HeLa: nuclear extract from HeLa cells grown in DMEM + 10% FBS.
Typical results obtained by using the NFkB p50 ELISA kit. Different amounts of nuclear extract as indicated were used for the assay. 293 TNFa: nuclear extract from HEK293 cells that were incu-bated overnight (16 hours) in DMEM containing 0.1% FBS followed by treatment with 20 ng/ml TNFa for 30 minutes. 293: nuclear extract from HEK293 cells that were incubated overnight (16 hours) in DMEM containing 0.1% FBS, untreated.
NFκB p50
R2 = 0.9915
R2 = 0.9976
0.0
0.2
0.4
0.6
0.8
1.0
1.2
1.4
1.6
1.8
0 2 4 6 8 10Nuclear Extract (µg)
293 TNFα
293
OD
450
ERα
Nuclear Extract (µg)
0.0
0.2
0.4
0.6
0.8
1.0
1.2
1.4
0 2 4 6 8 10
OD
450
HeLaMCF7
R2 = 0.9825
R2 = 0.9961
PPARα
0
0.5
1
1.5
2
0 10 20Nuclear Extract (µg)
OD
450
R2 = 0.9972
R2 = 0.4603
Cos-1Cos-1+PPARα+RXRA
Molecular Biology Products & Kits | Gene Regulation—Transcription Factors
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SP-1 ELISA KitThe SP-1 ELISA Kit quantifies SP-1 transcription factor activation.
Typical results obtained by using the SP-1 ELISA kit. Different amounts of recombinant human SP-1 protein as indicated were used for the assay.
Nuclear Extract (µg)
OD
450 R2 = 0.9993
SP-1
0.00
0.50
1.00
1.50
2.00
2.50
3.00
0.00 5.00 10.00 15.00 20.00 25.00
A450
TF Activation ELISA Kits continued...
HIF-1a ELISA KitThe HIF-1a ELISA Kit quantifies HIF-1a transcription factor activation.
Typical results obtained by using the HIF-1a ELISA kit. Different amounts of nuclear extract as indicated were used for the assay. Cos CoCl2: nuclear extract from Cos-1 cells treated overnight in DMEM + 0.1% FBS with 0.15 mM cobalt chloride. Cos: nuclear extract from Cos-1 cells incubated overnight in DMEM + 0.1% FBS, untreated.
p53 ELISA KitThe p53 ELISA Kit quantifies p53 transcription factor activation.
Typical results obtained by using the p53 ELISA kit. Different amounts of nuclear extract as indicated were used for the assay. MCF7: nuclear extract from MCF-7 cells incubated overnight in DMEM + 0.1% FBS, untreated. MCF7 H2O2: nuclear extract from MCF-7 cells incubated overnight in DMEM + 0.1% FBS, then treated with 0.2 mM H2O2 for 3 hours.
HIF-1α
Nuclear Extract (µg)
OD
450
R2 = 0.9986
R2 = 0.9876
Cos CoCl2Cos
0.0
0.2
0.4
0.6
0.8
1.0
1.2
1.4
1.6
0 2 4 6 8 10
Nuclear Extract (µg)O
D 4
50
R2 = 0.9974
R2 = 0.9969
p53
0.0
0.2
0.4
0.6
0.8
1.0
1.2
1.4
0 2 4 6 8 10
MCF7
MCF7 H2O2
AP-1 ELISA KitThe AP-1 ELISA Kit quantifies AP-1 transcription factor activation.
GR ELISA KitThe GR ELISA Kit quantifies GR transcription factor activation.
Typical results obtained by using the AP-1 ELISA kit. Different amounts of nuclear extract as indicated were used for the assay. 293 PMA: nuclear extract from HEK293 cells that were incubated over-night (16 hours) in DMEM containing 0.1% FBS, then treated with 10 ng/ml PMA for 4 hours. 293: nuclear extract from HEK293 cells that were incubated overnight (16 hours) in DMEM containing no FBS, untreated.
Typical results obtained by using the GR ELISA kit. Different amounts of nuclear extract as indicated were used for the assay. HeLa: nuclear extract from HeLa cells that were incubated overnight (16 hours) in DMEM + 0.1% FBS, untreated. HeLa Dex: nuclear extract from HeLa cells incubated overnight in DMEM + 0.1% FBS followed by treatment with 100 nM dexamethasone for 1 hour.
Nuclear Extract (µg)
OD
450
R2 = 0.9414
R2 = 0.9957
AP-1
293
293 PMA
0.0
0.2
0.4
0.6
0.8
1.0
1.2
0 2 4 6 8 10
Nuclear Extract (µg)
OD
450
R2 = 0.9899
R2 = 0.9865
GR
HeLa Dex
HeLa
0.0
0.2
0.4
0.6
0.8
1.0
1.2
1.4
1.6
0 2 4 6 8 10
STAT-1 ELISA KitThe STAT-1 ELISA Kit quantifies transcription factor activation, specifically that of STAT-1.
Typical results obtained by using the STAT-1 ELISA kit. Different amounts of nuclear extract as indicated were used for the assay. HeLa: nuclear extract from HeLa cells that were incubated overnight (16 hours) in DMEM containing no FBS, untreated. HeLa IFNγ: nuclear extract from HeLa cells incubated overnight in DMEM + 0.1% FBS followed by treatment with 5 ng/ml IFNγ for 2 hours.
Nuclear Extract (µg)
OD
450
R2 = 0.9891
R2 = 0.8107
HeLa
HeLa IFNg
STAT-1
0.0
0.2
0.4
0.6
0.8
1.0
1.2
0 2 4 6 8 10
Molecular Biology Products & Kits | Gene Regulation—Transcription Factors
109For bulk or alternate pack sizes, email us at [email protected].
TF Activation ELISA Kits continued...
MEF-2 ELISA KitThe MEF-2 ELISA Assay Kit quantifies MEF-2 tran-scription factor activation.
GATA-1 ELISA KitThe GATA-1 ELISA Kit quantifies transcription factor activation, specifically that of GATA-1.
Typical results obtained by using the MEF-2 ELISA kit. Different amounts of nuclear extract as indicated were used for the assay. U937: nuclear extract from U937 cells grown in DMEM + 10% FBS. C2C12: nuclear extract from C2C12 cells grown in DMEM + 10% FBS.
Typical results obtained by using the GATA-1 ELISA kit. Different amounts of recombinant human GATA-1 protein as indicat-ed were used for the assay.
Nuclear Extract (µg)
OD
450
R2 = 0.9955
R2 = 0.9945
U937
C2C12
MEF-2
0.0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
0 2 4 6 8 10Nuclear Extract (µg)
OD
450
R2 = 0.999A450
GATA-1
0.0
0.2
0.4
0.6
0.8
1.0
1.2
1.4
0 10 20 30 40
SMAD-2 ELISA Kit
HNF3b ELISA Kit
Typical results obtained by using the SMAD-2 ELISA kit. Different amounts of nuclear extract as indicated were used for the assay. HeLa: nuclear extract from HeLa cells incubated overnight in DMEM + 0.1% FBS, untreated. HeLa TGFb: nuclear extract from HeLa cells that were incubated overnight (16 hours) in DMEM con-taining 0.1% FBS followed by treatment with 10 ng/ml TGFb for 30 minutes.
Typical results obtained by using the HNF3b ELISA kit. Different amounts of nuclear extract as indicated were used for the assay. HepG2: nuclear extract from HepG2 cells that were grown in DMEM containing 10% FBS. HeLa: nuclear extract from HeLa cells grown in DMEM + 10% FBS.
SMAD-4 ELISA Kits
Typical results obtained by using the SMAD-4 ELISA kit. Different amounts of nuclear extract as indicated were used for the assay. HeLa: nuclear extract from HeLa cells incubated overnight in DMEM + 0.1% FBS, untreated. HeLa TGFb: nuclear extract from HeLa cells that were incubated overnight (16 hours) in DMEM con-taining 0.1% FBS followed by treatment with 10 ng/ml TGFb for 30 minutes.
Nuclear Extract (µg)
OD
450
R2 = 0.9878
R2 = 0.6832
HepG2
HeLa
HNF3β
0.0
0.2
0.4
0.6
0.8
1.0
1.2
1.4
1.6
0 5 10 15
Nuclear Extract (µg)
OD
450
R2 = 0.9967HeLa
HeLa TGFb
SMAD-2
0.0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
0 2 4 6 8 10
R2 = 0.9619
Nuclear Extract (µg)
OD
450
HeLa
HeLa TGFb R2 = 0.9936
R2 = 0.987
SMAD-4
0.0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0 2 4 6 8 10
Molecular Biology Products & Kits | Gene Regulation—Transcription Factors
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Typical results obtained by using the FKHR ELISA kit. Different amounts of nuclear extract as indicated were used for the assay. Raji: nuclear extract from Raji cells that were grown in RPMI-1640 containing 10% FBS. HeLa: nuclear extract from HeLa cells grown in DMEM + 10% FBS.
FKHR ELISA Kits
Nuclear Extract (µg)
OD
450
Raji
HeLa R2 = 0.9975
R2 = 0.954
FKHR
0.0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0 2 4 6 8 10
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TF Activation ELISA Kits continued...
E2F-1 ELISA KitThe E2F-1 ELISA Kit quantifies E2F-1 transcription factor activation.
Typical results obtained by using the E2F-1 ELISA kit. Different amounts of nuclear extract as indicated were used for the assay. HeLa: nuclear extract from HeLa cells that were incubated 3 days in DMEM containing no FBS. HeLa serum: nuclear extract from HeLa cells grown in DMEM + 10% FBS.
Nuclear Extract (µg)
OD
450
HeLa
HeLa serum
R2 = 0.9892
R2 = 0.9899
E2F-1
0.0
0.2
0.4
0.6
0.8
1.0
1.2
1.4
0 2 4 6 8 10
Product Size Prod. no.AP-1 Cold Probe 25 µl EK0504AP-1 ELISA Kit kit EK1040AP-1 ELISA Kit with Nuclear Extraction Kit kit EK1041Control HEK292 PMA Nuclear Extract 20 µl EK0505E2F-1 ELISA Kit kit EK2000E2F-1 ELISA Kit with Nuclear Extraction Kit kit EK2001ERa ELISA Kit kit EK1030ERa ELISA Kit with Nuclear Extraction Kit kit EK1031ER Control MCF7 Nuclear Extract 20 µl EK0503ERE Cold Probe 25 µl EK0502FKHR ELISA Kit kit EK1400FKHR ELISA Kit with Nuclear Extraction Kit kit EK1401GATA-1 Cold Probe 25 µl EK0520GATA-1 Control Recombinant Protein 20 µl EK0521GATA-1 ELISA Kit kit EK1100GATA-1 ELISA Kit with Nuclear Extraction Kit kit EK1101GR Cold Control Probe 25 µl EK0508GR Control Hela (DEX) Nuclear Extract 20 µl EK0509GR ELISA Kit kit EK1060GR ELISA Kit with Nuclear Extraction Kit kit EK1061HIF Cold Control Probe 25 µl EK0500HIF Control Cos1 Nuclear Extract 20 µl EK0501HIF-1a ELISA Kit kit EK1020HIF-1a ELISA Kit with Nuclear Extraction Kit kit EK1021HNF-3b ELISA Kit kit EK1800HNF-3b ELISA Kit with Nuclear Extraction Kit kit EK1801MEF-2 Cold Control Probe 25 µl EK0512MEF-2 Control Nuclear Extract 20 µl EK0513MEF-2 ELISA Kit kit EK1080MEF-2 ELISA Kit with Nuclear Extraction Kit kit EK1081NFkB c-Rel ELISA Kit kit EK4100
Product Size Prod. no.NFkB c-Rel ELISA Kit with Nuclear Extraction Kit kit EK4101NFkB Control Nuclear Extract 20 µl EK0523NFkB Cold Control Probe 25 µl EK0522NFkB p50 ELISA Kit kit EK1110NFkB p50 ELISA Kit with Nuclear Extraction Kit kit EK1111NFkB p52 ELISA Kit kit EK3100NFkB p52 ELISA Kit with Nuclear Extraction Kit kit EK3101NFkB p65 ELISA Kit kit EK1120NFkB p65 ELISA Kit with Nuclear Extraction Kit kit EK1121NFkB Rel-b ELISA Kit kit EK5100NFkB Rel-b ELISA Kit with Nuclear Extraction Kit kit EK5101p53 Cold Control Probe 25 µl EK0506p53 ELISA Kit kit EK1050p53 ELISA Kit with Nuclear Extraction Kit kit EK1051PPAR Cold Control Probe 25 µl EK0516PPARa Cold Control Probe 25 µl EK0518PPARa Control Cos1 Nuclear Extract 20 µl EK0519PPARa ELISA Kit kit EK1310PPARa ELISA Kit with Nuclear Extraction Kit kit EK1311SMAD-2 ELISA Kit kit EK1600SMAD-2 ELISA Kit with Nuclear Extraction Kit kit EK1601SMAD-4 ELISA Kit kit EK1500SMAD-4 ELISA Kit with Nuclear Extraction Kit kit EK1501SP-1 Cold Probe 25 µl EK0514SP-1 Control Recombinant Protein 20 µl EK0515SP-1 ELISA Kit kit EK1090SP-1 ELISA Kit with Nuclear Extraction Kit kit EK1091STAT-1 Cold Control Probe 25 µl EK0510STAT-1 Control MCF7 (IFNγ) 20 µl EK0511STAT-1 ELISA Kit kit EK1070STAT-1 ELISA Kit with Nuclear Extraction Kit kit EK1071
TF Activation ELISA Kits
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LRXXXX 10 µgPlease order using the product number identified on the chart below.
�� Physiologically relevant—contains cloned promoter region of specific genes �� Flexible—use for in vivo assays �� Quantitative—use luciferase activity to measure promoter activity �� Safe—no radioactivity required �� Simple—easily applied to high-throughput screening
With the Gene Promoter Reporter Vectors, you can quantify the promoter activity of 34 genes. The luciferase assay is a simple, straightforward, and very effective means of measuring promoter response in cells.
The actual length of the cloned promoter region dif-fers for each Gene Promoter Reporter Vector.
Gene Promoter Reporter Vectors
The Gene Promoter Reporter Vector is available for 34 different genes. The insert contains approximately 800 bp of 5' flanking region and 50-200 bp of untranslated region of exon 1 of the human genes listed below. This ~1 kb region is located upstream of the start codon of luciferase gene (Panel A.) The vector backbone contains an ampicillin-resistance gene which allows ampicillin selec-tion in E. coli, a pUC origin of replication for propagation in E. coli, and an f1 origin for single-stranded DNA production (Panel B).
The Gene Promoter Reporter Vectors in HeLa cells. HeLa cells were transfected with each of the vectors listed and then treated with/without phorbol myristate acetate (PMA). The luciferase activity was significantly enhanced with PMA treatment for TNFa, RB1, and JUNB Gene Promoter Vectors, indicating that these three gene promoters are activated by PMA in HeLa cells, while others are not.
Product Size Prod. no.14-3-3-sigma (SFN) Gene Promoter Reporter Vector 10 µg LR1009ABL1 Gene Promoter Reporter Vector 10 µg LR1001APC Gene Promoter Reporter Vector 10 µg LR1019ATF2 Gene Promoter Reporter Vector 10 µg LR1007BRCA1 Gene Promoter Reporter Vector 10 µg LR1002CFTR Gene Promoter Reporter Vector 10 µg LR1020CGPR (CALCR) Gene Promoter Reporter Vector 10 µg LR1003CIITA Gene Promoter Reporter Vector 10 µg LR1010COX2 (PTGS2) Gene Promoter Reporter Vector 10 µg LR1029ERa Gene Promoter Reporter Vector 10 µg LR1017FHIT Gene Promoter Reporter Vector 10 µg LR1032G6PD Gene Promoter Reporter Vector 10 µg LR1021GAGE1 Gene Promoter Reporter Vector 10 µg LR1004hMLH1 Gene Promoter Reporter Vector 10 µg LR1022HOXA2 Gene Promoter Reporter Vector 10 µg LR1030HOXA5 Gene Promoter Reporter Vector 10 µg LR1031IFNγ Gene Promoter Reporter Vector 10 µg LR1011
Product Size Prod. no.IGRP Gene Promoter Reporter Vector 10 µg LR1012IL1a Gene Promoter Reporter Vector 10 µg LR1013IL2 Gene Promoter Reporter Vector 10 µg LR1014IL4 Gene Promoter Reporter Vector 10 µg LR1023IRF7 Gene Promoter Reporter Vector 10 µg LR1005JUNB Gene Promoter Reporter Vector 10 µg LR1024MDR1 Gene Promoter Reporter Vector 10 µg LR1028MyoD Gene Promoter Reporter Vector 10 µg LR1025POMC Gene Promoter Reporter Vector 10 µg LR1006RB1 Gene Promoter Reporter Vector 10 µg LR1018STAT1 Gene Promoter Reporter Vector 10 µg LR1008SURVIVIN Gene Promoter Reporter Vector 10 µg LR1016THBS2 Gene Promoter Reporter Vector 10 µg LR1033TIMP3 Gene Promoter Reporter Vector 10 µg LR1034TNFa Gene Promoter Reporter Vector 10 µg LR1015VHL Gene Promoter Reporter Vector 10 µg LR1026v-myc Gene Promoter Reporter Vector 10 µg LR1027
Reporter vectors
A.
B.
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LRXXXX See belowPlease order using the product number identified in the chart below.
�� Quantify transactivation of transcription factor binding in vivo�� Target over 50 specific transcription factors�� Establish cell lines for cell-based assays and drug discovery
With Luciferase Reporter Vectors, you can easily measure the binding activities of transcription factors (TFs) in vivo. TF activation is measured by induced expression of firefly luciferase. This luciferase-based assay is simple, flexible, and rapid, generating quan-titative results in hours.
By simply transfecting a vector into cells, you can examine the effects of various stimuli on a given signaling pathway in vivo, such as CRE-luc for G-protein coupled receptors, NFkB-luc for TNFa, and STAT-luc for T cell antigen receptor. These vectors can be used to monitor disease-related TFs, such as PPAR for diabetes, NFkB for inflammatory disease, MEF-2 or MIF-1 for heart disease, and p53 for cancer.
Like our EMSA gel-shift kits, the Reporter Vectors are also ideal for validating the results obtained using Protein/DNA Arrays. But while EMSA is an in vitro assay, the Reporter Vectors allow you to monitor TF activity in living cells.
The Luciferase Reporter Vectors have been specially constructed to monitor transactivation of over 50 specific TFs. Each vector contains multiple repeats of a specific transcription factor binding element, or cis-element. Binding of a TF to its cis-element drives the expression of firefly luciferase. Light emitted from the chemical reaction is directly proportional to the amount of expressed luciferase, and thus the binding activity of the targeted TF.
Using Reporter Vectors is as simple as performing a standard luciferase assay, a procedure that takes just a few hours. Cells are transfected with a reporter vector, treated with corresponding stimuli, and then lysed to measure the luciferase activity.
By transfecting a reporter vector into cells along with vectors conferring resistance to hygromycin or neomycin, you can establish stable cell lines for use in cell-based assays.
Each Luciferase Reporter Vector comes with a Control Reporter Vector.
Luciferase Reporter Vectors
293, HeLa, or Cos-1 cells were transiently transfected with various Luciferase Reporter Vectors. The cells were then treated with inducers, the activity of each TF was then quantified by measur-ing luciferase activity.
Product Size Prod. no.TA-luc (Empty Control) Luciferase Reporter Vector 20 µg LR0000AP1(1) Luciferase Reporter Vector 10 µg LR0002AP1(2) Luciferase Reporter Vector 10 µg LR0003AP3 Luciferase Reporter Vector 10 µg LR0005AR Luciferase Reporter Vector 10 µg LR0007CRE(1) Luciferase Reporter Vector 10 µg LR0093E2F(1) Luciferase Reporter Vector 10 µg LR0016E2F(2) Luciferase Reporter Vector 10 µg LR0017E2F1(3) Luciferase Reporter Vector 10 µg LR0094ELK1 Luciferase Reporter Vector 10 µg LR0019ER Luciferase Reporter Vector 10 µg LR0020GAS / ISRE Luciferase Reporter Vector 10 µg LR0026GAS Luciferase Reporter Vector 10 µg LR0025GATA Luciferase Reporter Vector 10 µg LR0027GATA1 / GATA2 Luciferase Reporter Vector 10 µg LR0029GATA1 Luciferase Reporter Vector 10 µg LR0028GATA2 Luciferase Reporter Vector 10 µg LR0030GATA3 Luciferase Reporter Vector 10 µg LR0031GATA4 Luciferase Reporter Vector 10 µg LR0032GR Luciferase Reporter Vector 10 µg LR0033HIF1 Luciferase Reporter Vector 10 µg LR0128HSE Luciferase Reporter Vector 10 µg LR0038IRF1 Luciferase Reporter Vector 10 µg LR0039ISRE Luciferase Reporter Vector 10 µg LR0040
Product Size Prod. no.MEF1 Luciferase Reporter Vector 10 µg LR0043MEF2 Luciferase Reporter Vector 10 µg LR0044MEF3 Luciferase Reporter Vector 10 µg LR0045NF-AT Luciferase Reporter Vector 10 µg LR0050NFkB(1) Luciferase Reporter Vector 10 µg LR0051NFkB(2) Luciferase Reporter Vector 10 µg LR0052p53 Luciferase Reporter Vector 10 µg LR0057PR Luciferase Reporter Vector 10 µg LR0067RAR Luciferase Reporter Vector 10 µg LR0068RXR Luciferase Reporter Vector 10 µg LR0070RXR(2) Luciferase Reporter Vector 10 µg LR0114Smad Luciferase Reporter Vector 10 µg LR0116Smad3 / Smad4 Luciferase Reporter Vector 10 µg LR0115Sp1 Luciferase Reporter Vector 10 µg LR0071SP1(2) Luciferase Reporter Vector 10 µg LR0117SRE Luciferase Reporter Vector 10 µg LR0072SRF Luciferase Reporter Vector 10 µg LR0073STAT1 p84/p91 Luciferase Reporter Vector 10 µg LR0075STAT1(2) Luciferase Reporter Vector 10 µg LR0127STAT3(1) Luciferase Reporter Vector 10 µg LR0077STAT4 Luciferase Reporter Vector 10 µg LR0079VDR Luciferase Reporter Vector 10 µg LR0087YY1 Luciferase Reporter Vector 10 µg LR0090
Reporter vectors
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The Luciferase Reporter Vectors can be used to quantify induction in various cell lines. For the data shown above, 293, HeLa, and Cos-1 cells were transiently transfected with the indicated Luciferase Reporter Vector, in the presence or absence of an inducer molecule. The fold-induction was calculated for each vector relative to the unstimulated control. No induction by tested inducers was observed with the Negative Control Vectors.
Luciferase Reporter Vectors continued...
Molecular Biology Products & Kits | Gene Regulation—Transcription Factors
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MEXXXX 15 µgPlease order using the product number identified on the chart below.
�� Ready for transfection into cells for gene expression studies �� Fully-sequenced TF cDNA inserts
Transcription factor (TF) cDNA sequences are identi-cal to those on NCBI Refseq.
Mammalian Full-length TF Expression Vectors
Cloned human TF proteins are expressed at high levels from the constitutive CMV promoter. The SV40 polyA sequence ensures efficient processing of the 3' end of the mRNAs. The vector backbone contains an SV40 origin for replication in mammalian cells. A neomycin-resistance gene allows kanamycin selection in E. coli and neomycin selection in eukaryotic cells. The vector backbone also contains a pUC origin of replication for propagation in E. coli and an f1 origin for single-stranded DNA production.
pCMV-NFkB/p505748 bp
pUC ori
SV40 poly A
Not I
pSV40
Kan/Neor
NfkB/p50
pCMV
f1 ori
Xho I
HSV TK poly A
Product Size Prod. no.ATF1 CMV Expression Vector 15 µg ME0059ATF3 CMV Expression Vector 15 µg ME0055BTG2 CMV Expression Vector 15 µg ME0065CDX2 CMV Expression Vector 15 µg ME0060CITED1 CMV Expression Vector 15 µg ME0020E2F2 CMV Expression Vector 15 µg ME0010ERa CMV Expression Vector 15 µg ME0043FOS CMV Expression Vector 15 µg ME0044GATA1 CMV Expression Vector 15 µg ME0049GFI1 CMV Expression Vector 15 µg ME0066GR (NR3C1) CMV Expression Vector 15 µg ME0007HNF4γ CMV Expression Vector 15 µg ME0062IRF1 CMV Expression Vector 15 µg ME0014JUN CMV Expression Vector 15 µg ME0045MAFK CMV Expression Vector 15 µg ME0018MAX CMV Expression Vector 15 µg ME0019MYC CMV Expression Vector 15 µg ME0052 NF-ATC1 CMV Expression Vector 15 µg ME0046
Product Size Prod. no.NFkB p50 CMV Expression Vector 15 µg ME0005NFkB p65 (RelA) CMV Expression Vector 15 µg ME0006NF-YC CMV Expression Vector 15 µg ME0022NR0B1 CMV Expression Vector 15 µg ME0061NR1I2 CMV Expression Vector 15 µg ME0024NR1I3 CMV Expression Vector 15 µg ME0025p53 CMV Expression Vector 15 µg ME0047PBX1 CMV Expression Vector 15 µg ME0054PPARa CMV Expression Vector 15 µg ME0001PPARb (PPARD) CMV Expression Vector 15 µg ME0002PPARγ-1 CMV Expression Vector 15 µg ME0003PPARγ-2 CMV Expression Vector 15 µg ME0004PREB CMV Expression Vector 15 µg ME0029SMAD1 (MADH1) CMV Expression Vector 15 µg ME0071SRY CMV Expression Vector 15 µg ME0032STAT1 CMV Expression Vector 15 µg ME0033STAT5B CMV Expression Vector 15 µg ME0035USF1 CMV Expression Vector 15 µg ME0040
Expression vectors
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cAMP/Calcium Protein/DNA ArrayMA1500 kitcAMP/Calcium Protein/DNA Array RefillMA1600 kitCell Growth Protein/DNA ArrayMA1505 kitCell Growth Protein/DNA Array RefillMA1605 kitNuclear Receptor Protein/DNA ArrayMA1510 kitNuclear Receptor Protein/DNA Array RefillMA1610 kit
�� Focus on transcription factors involved in specific biological processes
We offer the Function-Specific Protein/DNA Arrays for function-specific analysis of eukaryotic TFs, allowing you to focus on factors involved in specific biological processes.
cAMP/Calcium Protein/DNA ArrayWith the cAMP/Calcium Protein/DNA Array, you can easily determine how the activities of cAMP- and calcium-regulated TFs change under different condi-tions. The array enables you to survey the activity of 20 TFs that are known to be regulated by cAMP and calcium.
Nuclear Receptor Protein/DNA ArrayThe Nuclear Receptor Protein/DNA Array is designed for researchers who are interested in how the activities of the lipophilic nuclear hormone receptor superfamily receptors—including steroid and thyroid hormones, active forms of vitamin A (retinoids), and
vitamin D—change under different conditions. With our Nuclear Receptor Protein/DNA Array, you can profile the activities of 15 nuclear receptors.
Cell Growth Protein/DNA ArrayThe Cell Growth Protein/DNA Array provides a good starting point for researchers who are looking to map the series of events leading from extracellular stimuli to gene transcription controlling growth and differen-tiation. This array allows screening for 20 TFs that are key players in cell growth and differentiation.
Kit components:Three array membranesHybridization and wash solutionsSpin column and buffersHRP chemiluminescent reagentsControl nuclear extract
Function-Specific Protein/DNA Arrays
TF-TF Interaction Array I*MA5010 kit TF-TF Interaction I RefillMA5110 kitTF-TF Interaction Array II*MA5011 kitTF-TF Interaction II RefillMA5111 kitTF-TF Interaction Array III*MA5012 kitTF-TF Interaction III RefillMA5112 kit
* See Protein/DNA Array chart on page 117 for cis-elements included on TF Interaction Array I, II and III.
�� Identify interactions between different transcription factors (TFs)�� Analyze transcription factor complex formation following different stimuli�� Monitor potential cross-talk between intracellular signaling cascades
TF-TF Interaction Arrays enable you to determine how a particular TF interacts with multiple other TFs—all in one experiment. Unlike traditional methods, such as co-immunoprecipitation and super-gel shift, this array-based method generates a global view of TF-TF interactions. The nonradioactive, all-in-one system offers the high sensitivity of HRP-based chemiluminescence detection.
Each consensus sequence immobilized on our TF-TF Interaction Array corresponds to a specific DNA-binding TF. We have 3 different arrays profiling a selection of transcription factors.
To prepare nuclear extracts for this procedure, please see the Nuclear Extraction Kit, PN AY2002 on page 102.
Here’s how it worksThe TF-TF Interaction Array, instead of directly measuring interactions between proteins, employs an exquisitely sensitive indirect approach. This technology is based on the fact that every TF binds to a specific DNA cis-element. The proteins present in a TF complex can be determined by isolating a complex by immunoprecipitation and monitoring which cis-elements bind to this complex—indicating which TF are present. In this way, you can quickly see how one TF interacts with multiple other TFs. Individual interactions can then be confirmed by co-immunoprecipitation or super-gel shift.
TF-TF Interaction Arrays
A. A431
The TF-TF Interaction Array I profiles interactions between multiple transcription factors. Nuclear extracts from A431 and Saos cells were incubated with Probe Mix I. The TF of interest, p53, was then immunoprecipitated using a p53-specific Ab, and the pro-tein-bound probes were isolated and hybridized to the TF-TF Interaction Array I revealing p53-specific interactions in A431 (Panel A), and no interactions in the p53 negative Saos2 cell line (Panel B).
B. Saos2
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Protein/DNA Array IMA1210 kitProtein/DNA Array I Refill KitMA1310 kitProtein/DNA Array IIMA1211 kitProtein/DNA Array II Refill KitMA1311 kitProtein/DNA Array IIIMA1212 kitProtein/DNA Array III Refill KitMA1312 kitProtein/DNA Array IVMA1213 kitProtein/DNA Array IV Refill KitMA1313 kitProtein/DNA Array VMA1214 kitProtein/DNA Array V Refill KitMA1314 kitCombo Protein/DNA ArrayMA1215 kitCombo Protein/DNA Refill KitMA1315 kit
�� Profile the activities of multiple transcription factors at once�� Monitor cross-talk between intracellular signaling cascades�� Characterize promoter regions of novel genes�� Determine, by high-throughput analysis, novel mechanisms of transcriptional regulation�� Greatly improved probe recovery system increases assay sensitivity
Protein/DNA Arrays simplify the functional analysis of eukaryotic transcription factors (TFs). Our array-based technology is a significant improvement over cumbersome gel mobility-shift assays, which permit the characterization of only a single TF at a time. With the Protein/DNA Arrays, you can profile the activities of multiple TFs simultaneously. The Protein/DNA Arrays can be used to study TF activation in a variety of biological processes, including cell proliferation, differentiation, transformation, and apoptosis.
Five versions of the Protein/DNA Array are currently available. All five arrays are spotted with unique, non-overlapping consensus-binding sequences corresponding to specific TFs, which were carefully selected after a survey of the literature. For your convenience we also offer a Combo version of the Protein/DNA Array spotted with all 345 cis-elements.
To prepare nuclear extracts for this procedure, please see the Nuclear Extraction Kit, PN AY2002 on page 102.
The new filter-based spin column system is a gel-free separation which makes probe isolation and recovery a quick and simple three-step process. This spin column separation procedure significantly increases the amount of probe recovered, providing a more sensitive assay that requires much less starting material. The reduced sample handling also helps minimize variability between experimental samples.
With increased probe recovery and ease of use, this system enables the whole experiment, from start to finish, to be completed in one and a half days. The added ease of sample preparation allows simultane-ous analysis of many more samples.
Kit components:Three array membranesHybridization and wash solutionsSpin column reagentsChemiluminescence reagentsControl nuclear extract
Refill Kits for reprobing array membranes can be purchased separately.
References:1. Jing, H. et al. (2004) J. Biol. Chem. 279,
55176–55186.2. Peterson, R. S. et al. (2004) J. Cell Biol. 166,
1081–1091.3. Barnhart, B. C. et al. (2004) EMBO J. 23,
3175–3185.4. Govindarajan, B. et al. (2003) J. Biol. Chem. 278,
9790–9795.
For some Transcription Factors, mutliple consensus sequences for the same TF have been published in the literature and, where appropriate, we have provided those variations available in our EMSA kits and arrays. For the TF’s with mutliple consensus sequences, they have been denoted with numbers in parenthesis, i.e. (1), (2), (3) etc. To obtain the consen-sus sequences and references for the TFs used in our arrays, please contact our Technical Support group.
Related products
Looking to profile transcription factors from a larger number of samples? Call our Technical Support team and ask about the Procarta Transcription Factor Plex Assays.
Protein/DNA Arrays
1. Mix biotin-labeled DNA binding probes with nuclear extract or cell lysate, this allows the formation of “DNA/Protein” complexes.
2. After incubation, these complexes are added to a spin col-umn which allows the separation of unbound probes.
3. After elution of the DNA/Protein com-plexes, they are denatured to liberate the free probe.
4. Hybridize the free probe to the mem-brane, which con-tains an array of Transcription Factor consensus binding sequences.
Here’s how it works
The Protein/DNA Arrays simultaneously detect changes in activities of multiple transcription factors. The array data was obtained using the Protein/DNA Array I. Nuclear extracts from untreated (panel A) or TNFa treated (panel B) HeLa cells were incubated with probe mix. The protein bound Probes were then isolated and hybridized to the Protein/DNA Array I membrane. Chemiluminescent images were collected using the FluorChem™ Imager (Alpha Innotech) revealing changes in protein binding activities of AP-1 and NFkB.
A. Untreated HeLa cells
B. TNFa treated HeLa cells
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Protein/DNA Arrays continued...
ADR-1
Afxh/Foxo-4
AhR/Arnt
ANG/IRE
Antioxidant RE
AP-3 (1)
AP-4 (1)
beta-RE
CdxA/NKX2
CEF-1
CEF-2
CETP/CRE
COUP-TF (1)
CREB-BP-1
c-Rel
DR-0
DR-1
DR-2
DR-3
DR-4
DR-5
E-47
E4BP-4 (1)
E4F/ ATF
Elk-1
EVI-1
FKHR (1)
FKHR (2)
Freac-2 (1)
Freac-2 (2)
Freac-4
Freac-7
GAG
GAS
GATA-1 (1)
GATA-1/2
GATA-2
GATA-3
GATA-4
GATA-6
GFI-1
HAS
HBS (1)
HBS (2)
HFH-1 (1)
HFH-2
HFH-3
HFH-8
HFS/HAS
HIF-1
HNF-3 (1)
HNF-3 (2)
HNF-4 (2)
HNF-4/COUP-TF
Ikaros
IRF-1/2
ISRE (1)
ISRE (2)
ISRE (3)
KKLF
L-III BP
MEF-2 (2)
MEF-2 (3)
MEF-3
MSP-1
MT-Box
MUSF-1
MZF-1 (1)
NF-Y (1)
Nkx-2.5
NPAS-2
NZF-3
ORE
p300
PARP
PAX-2 (1)
PAX-3
PAX-4
PAX-6 (1)
PAX-8
PPAR (2)
PPAR (3)
RREB (1)
RREB (2)
RSRFC4
SAA
Skn
SRY (1)
Tax/CREB
TCF/LEF
XBP-1 (1)
XRE (1)
ZIA
ZIB
ZIC
ZID
Protein/DNA Array II, TF-TF Interaction Array II, TF-Specific-TF Arrays II
ADD-1
AF-1
AFP-1
AIC/CBF
AML-1
AP-2/YY1
AREB-6
ARP
ATF (2)
ATF/CRE
CACC
CCAAT
CCAC
CD28RC (1)
CD28RC (2)
Cdx-2
CEA
CP1/CTF/CBTF
CPE
CREB-2
CSBP
CTCF
E12 (1)
EBP-40/45
EKLF (1)
EKLF (2)
ELF
H4TF
HEN-1
HFH-8/3/LUN
HiNF
HLF
HMG
HNF-1 (1)
HNF-1α
HOX4C
HOXD-8 (1)
HOXD-8 (2)
HOXD-8/9/10
HOXD-9/10
ICSBP
Isl-1
KPF-1
KTP-1
LCR-F1
LEF-1
LF-A1 (1)
LR-1
LyF-1 (1)
LyF-1 (2)
LyF-1 (3)
MAZ (1)
MAZ (2)
MTB-Zf
MTF (1)
MyoD
MyoG
MyTI
MZF-1 (2)
MZF-1 (3)
MZF-1 (4)
NF-4FA
NF-Gma (1)
NF-Gma (2)
NF-IL-2
NF-Y (2)
NRF-1
NRF-2
p53 (2)
p55
PEA3/HIP
PEBP
PEBP-2 (1)
PPUR (1)
PPUR (2)
PuF
Pur-1
PYR
RB
RFX-1/2/3 (1)
RFX-1/2/3 (2)
RIPE-3a1
SIF-1
SIF-2
SIF-3
SP-1 (2)
SRF/SAP
SRY (2)
TCE
TFE-3
Tf-LF
TGT-3
TIF-1
TREF-1/2
v-Maf
WT1 (1)
WT1 (2)
WT1 (3)
X2 BP
XBP-1/X2 BP
ZNF174
Protein/DNA Array III, TF-TF Interaction Array III, TF-Specific-TF Arrays III
AAF
ABF-1
ACF
ACP BP
AIC-2/3/4
ALF-1
α PAL
AP-2 (3)
AP-3 (2)
AP-4 (2)
apo A1 PR
ATF (1)
ATF (3)
beta-M-globin
factor B1
BZP
CBF (2)
CBFB
CEBP (2)
CEBP (3)
CEBP (4)
CEBP (5)
c-Ets-1
c-fos BP
c-Myb (2)
c-Myc
COUP-TF (2)
CP-1
CP-1B
CREB (2)
CYP1A1
DE-1
Dioxin Receptor
E12 (3)
E12/E47
E2
E2F-1 (2)
E4BP-4 (2)
EBP-80
EBP-C2/
NF-muE3/YEB3
EGF BP
EGR-1 (2)
EGR-2 (1)
EIL-1/2/3
ETF
ETV4/E1A-F
Fra-1/JUN
Gastrin EGF RE
GATA-1 (2)
GBF-1/2/3/HY5
GKLF
H4TF-1
HFH-1 (2)
HFH-11β/11α
HiNF/D3
HiNF-A
HiNF-B/H1TF1
HNF-1 (2)
HNF-1 a/b/c
HNF-4α2/1
HOXA-4
HSF/HTF
HSF-2 distal
HSF-3 proximal
IFI-56KBP
IL6-RE-BP
IR-4/EBV BP
ISGF
kBF-α
lactoferrin BP
LF-A1 (2)
LF-A2
LF-B2
LH2/Lim1
LSF
LXRE-1
MASH-1
MBP-1 (1)
MBP-1 (2)
Protein/DNA Array IV
mck PR E1
MDBP (1)
MDBP (2)
MEF-2a
Mfh-1
MHC gene PR
W Box
msx-1/2/3
Myb (2)
myc/CF1
myc-PRF
NCAM-BP
NF-1 (2)
NF-1/L
NF-A
NF-A3
NFAT-1 (2)
NF-Atp
NF-Atx
NF-E2 (2)
NF-E6/CP1
NFκB (2)
NFκB (3)
N-ras BP
OCT (2)
OCT (3)
ODC
p53 (3)
PAX-1
PAX-2 (2)
PAX-5 (2)
PAX-6 (2)
PBGD BP
PCF
PEBP-2 (2)
PEPCK PR GR
Pit-1 (2)
Pit-1 (3)
PO-B
PRDI-BFc
PRDII-BF1
PREB
PTF-1
PTF-1β
PU.1
PUR
RAR/RXR/CF1/
MB67
RFX-1/2/3 (3)
RORE
RVF
SAP-1b
Snail
SPERM-1
SRF (1)
SRF (2)
SSAP
Stat-1/3
Stat-5b
Surf-2 (2)
T3R
Tat
TFE-3L
TFEB
Thy-1 BP
tPA BP
Transferrin BP
TTF-1 (1)
TTF-1 (2)
TxREF/NF-lll
URE
WAP BP
XBP-1 (2)
XRE (2)
YB-1
Protein/DNA Array V
AP-1 (1)
AP-1 (2)
AP-2 (1)
AP-2 (2)
AR
Brn-3
CBF (1)
CDP
CEBP (1)
c-Myb (1)
CREB (1)
E2F-1 (1)
EGR-1 (1)
ER
Ets
Ets-1/PEA-3
FAST-1
GAS/ISRE
GATA
GR/PR
HNF-4 (1)
HSE
IRF-1
MEF-1
MEF-2 (1)
MRE
Myc-Max
NF-1 (1)
NFAT-1 (1)
NF-E1/YY1
NF-E2 (1)
NFκB (1)
OCT (1)
p53 (1)
PAX-5 (1)
Pbx-1
Pit-1 (1)
PPAR (1)
PRE
RAR/DR-5
RXR/DR-1
SIE
SMAD/SBE
SMAD-3/4
Sp-1 (1)
SRE
Stat-1
Stat-3
Stat-4
Stat-5
Stat-6
TFIID
TR
TR/DR-4
USF-1 (1)
VDR/DR-3
Protein/DNA Array I, TF-TF Interaction Array I, TF-Specific-TF Arrays I
Protein/DNA Array Content
Molecular Biology Products & Kits | Gene Regulation—Transcription Factors
118 888-362-2447 | 216-765-5000 | usb.affymetrix.com
NFkB p50-TF Interaction Array IMA5051 kitNFkB p50-TF Interaction Array IIMA5052 kitNFkB p50-TF Interaction Array IIIMA5053 kitNFkB p65-TF Interaction Array IMA5054 kitNFkB p65-TF Interaction Array IIMA5055 kitNFkB p65-TF Interaction Array IIIMA5056 kitPPARa-TF Interaction Array IMA5041 kitPPARa-TF Interaction Array IIMA5042 kitPPARa-TF Interaction Array IIIMA5043 kitPPARγ-TF Interaction Array IMA5044 kitPPARγ-TF Interaction Array IIMA5045 kitPPARγ-TF Interaction Array IIIMA5046 kit
�� A line of arrays for analysis of specific transcription factor complexes, focusing on interactions with NFkB and PPAR.
NFkB-TF Interaction Array NFkB-TF Interaction Array profiles interactions between NFkB and multiple TFs in a single hybrid-ization experiment and allows you to compare the interaction patterns under different conditions. Un-like traditional methods, this array-based technique generates a global view of interactions between NFkB and other proteins.
We currently offer three versions of the NFkB p50− and NFkB p65−TF Interaction Arrays, respectively, profiling a total of 244 different Transcription Factors (TFs). Each array is spotted with TF consensus-DNA binding sequences.
Here’s how they workThe TF-Specific-TF Interaction Arrays employ a pro-prietary, patent-pending technology that facilitates screening multiple interactions at once. The method is essentially the same as that used with the TF-TF Interaction Arrays, and the TF specific antibody is included for convenience.
References:1. Van Le, T. S., et al. (2004) Clin. Cancer Res. 10(4),
1384-1391.2. Hewetson, A., et al. (2003) J. Biol. Chem. 278(41),
40177-40185.
NFkB-TF and PPAR-TF Interaction Arrays
Nuclear extracts from untreated or TNFa-treated HeLa cells were incubated with the NFkB Probe mix. Immunoprecipitation with a NFkB-specific antibody was performed, and the protein-bound probes were then isolated and hybridized to the NFkB-TF Interaction Array membrane. Chemiluminescent images were collected using a FluorChem imager (Alpha Innotech), revealing changes in TF binding to NFkB upon TNFa stimulation.
Nuclear extracts from HeLa cells were incubated with the PPARa or PPARγ Probe Mix. Immunoprecipitation with the corre-sponding PPAR-specific antibody was performed, and the protein-bound probes were then isolated and hybridized to the PPARa or PPARγ-TF Interaction Array membrane. Chemiluminescent images were collected using a FluorChem imager (Alpha Innotech), revealing differences in PPARa- and PPARγ-interacting transcription factors in HeLa cells.
PPAR-TF Interaction ArrayPPAR-TF Interaction Array profiles interactions be-tween specific PPARs and multiple TFs. We currently offer three versions of the PPARa− and PPARγ−TF Interaction Arrays, respectively.
HeLa HeLa-TNFa
PPARa with HeLa extract PPARγ with HeLa extract
Molecular Biology Products & Kits | Gene Regulation—Transcription Factors
119For bulk or alternate pack sizes, email us at [email protected].
See chart on page 122 for full listing of Reporter Stable Cell Lines
�� Perform cell-based assays without transfection�� Screen compounds that inhibit or activate pathway components�� Monitor biochemical changes in NFkB, STAT, NFAT, CREB, AP-1, and SRF pathways
We offer NFkB, STAT-1, STAT-3, GR, NFAT, CREB, AP-1, SRF, TAD Kinase, and HIF Reporter Stable Cell Lines for high-throughput analysis of in vivo drug efficacy and specificity. These cells also provide a reproducible, ready-to-use system for performing cell-based assays. Among other applications, they can be used to evaluate uncharacterized growth factors, extracellular stimuli, and upstream events in the NFkB, STAT, NFAT, CREB, AP-1, SRF, TAD, and HIF signaling pathways. We currently offer NFkB Reporter Stable Cell Lines in five different cell lines: human A549, HeLa, 293, NIH3T3, and C2C12 cells. STAT-1 Reporter Stable Cell Lines are available in HeLa cells, NFAT in HeLa and K562, CREB in CHO and 293 cells, AP-1 in 293 cells, SRF in HeLa, and HIF in NIH3T3 cells.
All of the Reporter Stable Cell Lines maintain chromosomal integration of a luciferase reporter construct regulated by multiple copies of the transcription factor response element. The clonal cell lines are obtained by co-transfection of the luciferase reporter construct and pHyg followed by hygromycin selection.
NFkB p50 knockdownWe have established the NFkB p50 knockdown stable HeLa cells using a functionally validated shRNA expression vector. This cell line stably expresses siRNA specific for NFkB p50, and effectively knocks down the gene transactivation activity of NFkB. It is a powerful tool to identify novel NFkB target genes, and to analyze the interaction of other signaling proteins with NFkB. This stable cell line enables identification of downstream target genes of NFkB and allows the analysis of the cross-talk between NFkB and other signaling proteins.
NFkB Reporter Stable A549, HeLa, 293, NIH3T3 and C2C12 Cell LinesNuclear Factor kappa B (NFkB) is a member of the rel family of transcription factors. NFkB plays a key role in the regulation of inflammatory response, apoptosis, and tumorigenesis. NFkB is activated by a wide variety of different stimuli, including proinflammatory cytokines, oxidant free radicals, inhaled particles, ultraviolet radiation, and bacterial or viral products.
We have established NFkB reporter stable cells in a number of popular cell lines suitable for specific applications in the study of NFkB. They include 293 human embryonic kidney cells, HeLa human cervical epithelial cells, NIH3T3 mouse fibroblast cells, A549 human lung carcinoma cells, and C2C12 mouse myoblast cells. All these cell lines have been used extensively for the study of NFkB signaling due to their robust response. The 293 and HeLa cells are highly transfectable human cell lines whereas murine NIH3T3 cells are easy to maintain and transfect. In addition, A549 and C2C12 cells are useful models for studying the role of NFkB in lung carcinogenesis and muscle differentiation, respectively.
Reporter Stable Cell Lines
These cells have been treated with TNFa to induce lucifer-ase activity.
NFkB responsive luciferase reporter construct was trans-fected into the p50 knockdown stable cells or parental HeLa cells.
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Luci
fera
se A
ctiv
ity
18,000
16,000
14,000
12,000
10,000
8,000
6,000
4,000
2,000
0
Parentalp50 KD
control
TNFα
Molecular Biology Products & Kits | Gene Regulation—Stable Cell Lines
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Reporter Stable Cell Lines continued...
STAT-1 Reporter Stable HeLa Cell LineSTAT-1 is activated upon binding by IFN a, b, or γ; or IL-6 to its cognate receptor. Once STAT-1 is phosphorylated by JAKs, it forms a homodimer, or a heterodimer with STAT-2, and translocates to the nucleus. Once in the nucleus, the dimer binds to the GAS sequence (IFNγ activation site) in the promoter region of target genes and induces transcription.
These cells have been treated with IFNγ to induce luciferase activity.
AP-1 Reporter Stable 293 Cell LineThe activator protein 1 (AP-1) transcription factor is a dimeric complex that comprises members of the JUN, FOS, ATF, and MAF protein families with FOS and JUN being the most common of these proteins in mammalian cells. AP-1 activity is induced by growth factors, cytokines, and oncoproteins. Upon activation, AP-1 binds to the TPA Responsive ele-ment (TRE) and induces transcription of a variety of genes involved in multiple cellular functions such as proliferation, survival, differentiation and transforma-tion. The regulation of AP-1 activation is associated with the expression of genes important to cell differentiation programs.
CREB Reporter Stable CHO and 293 Cell LinesThe cyclic AMP response element (CRE)-binding protein (CREB) is one of the best-understood phos-phorylation-dependent transcription factors. Upon stimulation of cellular G-protein-coupled receptors and growth factor receptors, CREB is phosphory-lated by PKA, in a cAMP-dependent fashion. CREB can also be phosphorylated by a variety of other kinases in response to mitogen, calcium, and stress dependent signals. When phosphorylated on Ser133, CREB recruits transcriptional co- activators that induce transcription of a variety of intermediate early response genes. CREB is central to cellular processes such as growth factor-dependent cell proliferation and survival, and glucose homeostasis in learning and memory.
NFAT Reporter Stable HeLa and K562 Cell LinesNFAT transcription factors are activated by the calcium- and calmodulin-dependent serine phosphatase, calcineurin. Once dephosphorylated, NFAT translocates to the nucleus, where it acts synergistically with a wide variety of other transcription factors, such as AP-1, GATA, EGR, OCT, MEF-2, PPARγ, and HDACs. NFAT is subsequently phosphorylated and inactivated by a variety of kinases, such as GSK3, p38-MAPK, and PKA. The regulation of NFAT activation is associated with the expression of genes important to cell differentiation programs.
These cells have been tested with PMA to induce luciferase activity.
These cells have been tested with Forskolin to induce lucif-erase activity.
These cells have been treated with PMA and a calcium ionophore to induce luciferase activity.
Molecular Biology Products & Kits | Gene Regulation—Stable Cell Lines
121For bulk or alternate pack sizes, email us at [email protected].
Glucocorticoid Receptor Reporter 293 and HeLa Stable Cell LinesGlucocorticoids, a major subclass of steroid hormones, modulate a large number of metabolic, cardiovascular, immune, and behavioral functions. The intracellular effects of glucocorticoids are mediated by the glucocorticoid receptor (GR), a 94-kDa intracellular protein belonging to the phylogenetically conserved nuclear hormone receptor superfamily. In the absence of glucocorti-coid, the GR is maintained predominantly in the cytoplasm as part of an inactive multiprotein complex that consists of the receptor, two Hsp90 molecules, one molecule each of Hsp70 and Hsp56, and an immunophilin of the FK506- and rapamycin-binding class. When glucocorticoid binds to GR, the receptor undergoes a change in conformation, dissociates from regulatory heat shock proteins, and is hyperphosphorylated. The activated receptor rapidly translocates to the cell nucleus, where it is able to initiate transcriptional regulation.
In the nucleus, hormone-activated GR regulates transcription. During transcriptional regulation, the GR binds as a homodimer directly to short, palindromically arranged DNA sequences located in the promoter regions of glucocorticoid-responsive genes. Binding to these sequences, known as glucocorticoid response elements, leads to transcriptional induction or repression of target genes.
Induction of Luciferase Activity by Different Stimuli.
TAD-Kinase Reporter Stable Cell LineA previous system, the TAD in vivo Kinase Assay Kit, was developed to detect phosphorylation of the TAD of a specific TF. Now, to make this assay system more user-friendly, we have stably integrated the pGAL4-Luciferase vector into HeLa cells. Since only one vector, pGAL4-TAD, is needed for transfection into these cells, the sensitivity of this assay is dramatically increased. Upon stimulation, activated kinases will mediate modification of TADs leading to induction of the luciferase gene. Alternatively, kinases can be over-expressed via co-transfection with pGAL4-TAD and the activity of the kinase can be measured in vivo. Furthermore, the cells can be used in a mam-malian two-hybrid system for detecting protein-protein interaction.
These cells have been tested with Cobalt Chloride to induce luciferase activity.
SRF Reporter Stable HeLa Cell LineSerum response factor (SRF) is a widely expressed transcription factor involved in the expression of genes linked to cellular growth and muscle differen-tiation. Its target genes include cellular immediate early genes such as c-fos and structural cytoskeletal proteins, such as actins, myosins, tropomyosin, and vinculin. Serum-induced transcriptional activation of immediate early genes involves the formation of a ternary complex of SRF with a 62 kDa ternary complex factor (TCF) which binds to the serum response element (SRE). SRF is also involved in vascular smooth muscle differentiation by regulating the expression of several contractile and cytoskeletal genes. It is widely accepted that various diseases subvert normal smooth muscle cells to one of growth and excess matrix production.
These cells have been tested with Serum + PMA to induce luciferase activity.
HIF NIH3T3 Reporter Stable Cell LineHypoxia inducible factor (HIF) is a heterodimeric transcription factor composed of two basic helix-loop-helix proteins—HIF and HIF-1‚ (or Arnt). The HIFa/b dimer binds to hypoxia-response element and activates the transcription of many
genes that code for proteins that are involved in angiogenesis, glucose metabolism, cell proliferation/survival and invasion/metastasis, such as VEGF, FLT-1, EPO, and Leptin. Studies have shown the association of aberrant HIF activities with human diseases such as cancer and heart disease.
We have established HIF reporter stable cells in murine fibroblast NIH3T3 cells suitable for various applications in the research of HIF including the study of drug potency and the evaluation of novel inhibitor or activator proteins.
These cells have been tested with Cobalt Chloride to induce luciferase activity.
GRa CHO GFP Reporter Stable Cell LineThe CHO/ GFP-GRa cell line is designed for monitor-ing the activity of glucocorticoid receptor GRa in cell-based assays. The CHO/ GFP-GRa cell line was obtained by co-transfection of an expression vector for a fusion protein of TurboGFP (Evrogen PN FP511) and human GRa, as well as pHyg into Chinese ham-ster ovary (CHO) cells. Cells were then grown in the presence hygromycin. Single cells with strong green fluorescence were selected by flow cytometry, and allowed to expand. These cells constitutively express the GFP- GRa fusion protein.
NFkB CHO GFP Reporter Stable Cell LineThe CHO/ GFP-NFkBp65 cell line is designed for monitoring the activity of NFkB transcription factor in cell-based assays. The CHO/ GFP-NFkBp65 cell line was obtained by co-transfection of an expres-sion vector for a fusion protein of TurboGFP (Evrogen PN FP511) and human NFkBp65, as well as pHyg into Chinese hamster ovary (CHO) cells. Cells were then grown in the presence hygromycin. Single cells with strong green fluorescence were selected by flow cytometry, and allowed to expand. These cells constitutively express the GFP-NFkBp65 fusion protein.
Reporter Stable Cell Lines continued...
a γ
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Molecular Biology Products & Kits | Gene Regulation—Stable Cell Lines
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Reporter Stable Cell Lines continued...
Product Size Prod. no.AP-1 293 Reporter Stable Cell Line vial RC0006CHO/ GFP-GRa Stable Cell Line vial RC2002CHO/ GFP-NFkBp65 Stable Cell Line vial RC2001CREB 293 Reporter Stable Cell Line vial RC0007CREB CHO Reporter Stable Cell Line vial RC0008GR 293 Reporter Stable Cell Line vial RC0018GR HeLa Reporter Stable Cell Line vial RC0019HIF NIH3T3 Reporter Stable Cell Line vial RC0017NFAT HeLa Reporter Stable Cell Line vial RC0005NFAT K562 Reporter Stable Cell Line vial RC0009
Product Size Prod. no.NFkB 293 Reporter Stable Cell Line vial RC0014NFkB A549 Reporter Stable Cell Line vial RC0002NFkB C2C12 Reporter Stable Cell Line vial RC0016NFkB HeLa Reporter Stable Cell Line vial RC0013NFkB NIH3T3 Reporter Stable Cell Line vial RC0015NFkB p50 Knockdown HeLa Stable Cell Line vial RC5001SRF HeLa Reporter Stable Cell Line vial RC0012STAT-1 HeLa Reporter Stable Cell Line vial RC0004TAD-Kinase Reporter Stable Cell Line vial RC1001
Related products
Reporter stable cell lines can be stimulated with a variety of molecules. Some of these (cytokines, drugs, etc.) typically act at the cell surface, while others (siRNA, peptides) require transfection inside the cell.
By using the MPG amphipathic peptide rather than lipid mediated transfection, our DeliverX family of transfection reagents delivers siRNA or peptides to both cultured and primary cell lines with high efficiency and very low toxicity. See pages 98-100.
Reporter Stable Cell Lines
Molecular Biology Products & Kits | Gene Regulation—Stable Cell Lines
123For bulk or alternate pack sizes, email us at [email protected].
Molecular Biology Products & Kits | Next Generation Sequencing Library Prep
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79900 10 reactions 20 reactions
Generate high quality next generation sequencing (NGS) libraries suitable for sequencing on Illumina platforms in less time with Prep2Seq DNA Library Prep Kit.
DNA library prep is critical to the precision and accuracy of NGS. The Prep2Seq DNA Library Prep Kit comes with all of the reagents necessary for end repair, A-tailing, and ligation, and can be completed in less than two hours.
Our kit is fast, efficient, and optimized. It efficiently converts 3’ and 5’ overhangs into 3’ A-overhang ends for ligation to T-tailed DNA adapters and reduces the potential for bias with our superior A-tailing and ligation steps. We offer significant improvement in the quality of sequence-ready libraries by eliminating chimeras and reducing side reaction fragments.
The Prep2Seq DNA Library Prep Kit combines end re-pair and A-tailing steps, decreasing the total number of steps to achieve quality NGS library preparation. Only one cleanup step is required to attain peak performance on the Illumina sequencing platform and only 100 ng of input is needed.
Full kit offering�� Single kit that includes all reagents for sample prep in a slimmed down protocol.�� 24 bar code adapter sequences compatible with Illumina technology are sold separately:
Prep2Seq Multiplex Oligo Adapters for Illumina - Index Adapter Set I–adapter sequences 1-12 - Index Adapter Set II–adapter sequences 13-27
Time savings with a more efficient method for NGS sample prep�� Improved A-tailing reaction with proprietary enzymes, cloning know-how, and optimized buffers.�� By combining the end repair and A-tailing steps, only one cleanup step is needed compared to the two or three steps in most other kits (saving up to two and a half hours in the lab).
The Prep2Seq DNA Library Prep Kit offers the highest adapter ligation efficiency available. By combining end repair and A-tailing, the Prep2Seq DNALibrary Prep Kit is able to reduce total NGS library prep time to 110 minutes and limit bead cleanup to a single step.
Rigorous comparison studies to competitor kits show that the Prep2Seq DNA Library Prep Kit generates impressive sequencing data.
Prep2Seq™ DNA Library Prep Kit for Illumina platform
Illumina TruSeq™ Nano
140 min
NEB Next® Ultra
65 min
USB Prep2Seq™
40 minHands-on Lab Time
Walk-Away Time
USB - 110 total min
NEB - 155 total min
Illumina - 210 total minIllumina TruSeq Nano
70 min
NEB Next Ultra
90 min
USB Prep2Seq
70 min
Time Comparison
from sheared DNA to size selection
Table 1. Validated performance of the Prep2Seq DNA Library Prep Kit compared to competition.
USB® Prep2Seq™ Illumina® TruSeq™ NEBNext® Ultra™
Total Reads 170,507,429 171,139,925 176,073,965
Mapped reads to a single locus 142,223,440 143,567,898 147,688,157
% Mapped reads to a single locus 83.41% 83.89% 83.88%
Mapped reads to multiple loci 12,270,818 11,794,099 12,266,301
% Mapped reads to multiple loci 7.20% 6.89% 6.97%
Unmapped reads 16,013,171 15,777,928 16,119,507
% Unmapped reads 9.39% 9.22% 9.15%
The HiSeq® 2500 raw reads were aligned to the human genome hg19 release using Bowtie, allowing 2 mismatches for the entire read length. Mapped reads aligning to a single locus, mapped reads aligning to multiple loci, as well as unmapped reads from the 3 libraries are summarized in Table 1.
Table 2. GC content present in mapped and unmapped reads for the 3 library prep methods.
USB® Prep2Seq™ Illumina® TruSeq™ NEBNext® Ultra™
% GC: Mapped reads to a single locus 42.27% 40.01% 41.10%
% GC: Mapped reads to multiple loci 43.53% 41.65% 42.55%
% GC: Unmapped reads 40.90% 38.79% 40.64%
In order to determine if the end-repair and adaptor ligation may have contributed to some library bias we compared the overall GC content in mapped and unmapped reads for each of the 3 kits. As shown in Table 2, no substantial difference in GC content was introduced during the preparation methods of the 3 kits.
Time Comparison
from sheared DNA to size selection
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Molecular Biology Products & Kits | Next Generation Sequencing Library Prep
Prep2Seq™ DNA Library Prep Kit for Illumina platform continued...
79800 1 KT Set 1(Contains adapter sequences 1-12) 2 KT Set 2(Contains adapter sequences 13-27)
Prep2Seq Multiplex Oligo Adapters allow sample multiplexing in NGS reactions. These are HPLC purified adapter duplex sets that are bar coded and designed for multiplex sample preparation for Next Generation Sequencing on Illumina platforms.
Set 1 components:12 multiplex adapters that correspond to Illumina's
adapters 1-12.20 reactions per adapter Each adapter vial contains 4.8 nmol, 80 μl.
Set 2 components:12 multiplex adapters that correspond to Illumina's
adapters 13-16, 18-23, 25, and 27. 20 reactions per adapterEach adapter vial contains 4.8 nmol, 80 μl.
Prep2Seq Multiplex Oligo Adapters for Illumina platform
TOTAL kit time
Workflow Time Comparison
from sheared DNA to size selection
Hands-on Lab Time-
Walk-Away Lab Time-
VS.
TTTTTTOOOTALTTtt tttttimimimimime
WaWW lk-Away
min Ligation set up
Illumina TruSeq Nano
NEB Next Ultra
USB Prep2Seq 5
min Ligation set up
30min Incubation
40min Incubation
60min Incubation 30min
Incubation55min
Clean up
5 min
End repair set up
30min Incubation
35 min
Clean up
5 30min Incubation
5
5 10min Incubation
90min Clean up
30min Clean up
5 min End repair/ A-tail set up
min A-tail set up
5 min End repair/ A-tail set up
210min TOTAL kit time
155min TOTAL kit time
110min TOTAL kit time
min Ligation set up
The Prep2Seq DNA Library Prep Kit ensures efficiency of ligation to T-tailed adapters by optimizing the A-tailing step. This enzymology focus on A-tailing ensures that end bias is reduced, and the formation of blunt- and G-tailed ends is minimized. Most enzymes used for A-tailing are inefficient and result in a large amount of blunt-end DNA that can ligate as concatemers, forming chimeric false joints and compromising sequencing data. Effective A-tailing is crucial in preparing efficient DNA adapter libraries that are ready for sequencing.
Also available separately are barcoded adapter sets that enable multiplexing of up to 24 samples. Prep2Seq Multiplex Oligo adapters for Illumina systems are available in kits of 1-12 and 13-27 duplex (79800 1 KT and 79800 2 KT).
Kit components:This kit is designed for input sheared genomic DNA amounts from 100 ng to 1 μg and consists of 3 tubes: Prep2Seq NGS Enzyme Mix, Prep2Seq NGS 2X Buffer, and Prep2Seq NGS High Concentration
Ligase. A streamlined workflow, straightforward pro-tocol, and high performing kit ensures quality library prep for your Illumina sequencing runs.
Per 10 reactions: Prep2Seq NGS Enzyme Mix, 40 μl Prep2Seq NGS 2X Buffer, 250 μl Prep2Seq NGS High Concentration Ligase, 40 μl
Shipping and storage:Shipped on dry ice. Store at -20°C.
WorkflowTime Comparison
from sheared DNA to size selection
125For bulk or alternate pack sizes, email us at [email protected].
78400 25 reactions78410 100 reactions
Description:The Ligate-IT Rapid Ligation Kit provides high-quality reagents for fast and easy ligation.
Quick and efficientLigation reactions are complete following room temperature incubation in just 5 minutes for cohesive-ends and 10 minutes for blunt-ends (Figs. 1 - 2). Transform competent cells immediately or store reactions at -20°C for later transformation.
ConvenientLigate-IT Rapid Ligation Kit includes T4 DNA Ligase, Nuclease-Free Water, and a specially formulated 5X Reaction Buffer which enhances the activity of T4 DNA Ligase, especially on blunt-end fragments. All of the components are in a ready-to-use format and do not require any additional reagents, such as ATP.
FlexibleThe kit can be used for a variety of ligation procedures such as conventional vector cloning, TA cloning, linker or adaptor ligation, and library construction.
Functional test:Cohesive- and blunt-end ligations of a 1200 bp insert into pUC19 yields ≥ 1 x 106 transformants per microgram of linearized, dephosphorylated vector.
Kit components: 25 Reaction 100 Reaction Pack Size Pack Size
Ligate-IT T4 DNA Ligase 25 µl 100 µl5X Ligate-IT Reaction Buffer 100 µl 400 µlWater, Nuclease-Free 1 ml 2 x 1 ml
Shipping and storage:Shipped on dry ice. Store at -20°C.
Reference:1. Sambrook, J. and Russell, D. W. (2001) “Molecular
Cloning: A Laboratory Manual,” Cold Spring Harbor Laboratory Press, 1.84-1.125.
Related products:
FideliTaq DNA Polymerase71180 50 units 250 units 1,000 units 5 x 250 units 5,000 units
HotStart-IT FideliTaq DNA Polymerase71155 50 units 250 units 1,000 units 5,000 units
OptiKinase78334X 500 units78334Y 1,000 units78334Z 2,500 units
Shrimp Alkaline Phosphatase78390 500 units 1,000 units 5,000 units
SuperSAP Shrimp Alkaline Phosphatase75592 50 reactions
Ligate-IT Rapid Ligation Kit
Fig. 1. Rapid ligation of cohesive-end and blunt-end frag-ments. Lambda DNA was cut with Aat II (cohesive-end fragments) or EcoR V (blunt-end fragments) and purified. In each lane, 500 ng of cut lambda DNA were ligated with the Ligate-IT Rapid Ligation Kit for the indicated times (in minutes) and then loaded onto a 1% agarose gel. M are the DNA marker lanes. Results demonstrate fast and efficient ligation of both cohesive-end and blunt-end fragments in just 5 minutes.
Fig. 2. Ligate-IT Rapid Ligation Kit transformation efficiency of cohesive-end and blunt-end ligations. A modified pUC19 vector was cut to generate cohesive-ends (Aat II and EcoO109 I) and blunt-ends (Stu I). Both vectors were dephosphorylated prior to liga-tion. The lac repressor (lacI) from E. coli was cloned into the modified pUC19 vector with matching restriction sites for both cohesive-end and blunt-end ligations. Transformation efficiencies were determined by counting white colonies and are expressed in colony forming units per microgram of DNA used during transformation (cfu/µg). The uncut, supercoiled control vector was pUC19-lacI. Results indi-cate at least 106 transformants are obtained for both cohesive-end and blunt-end ligations when using competent cells that yield >108 cfu/µg.
Molecular Biology Products & Kits | Cloning
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Molecular Biology Products & Kits | Mutagenesis
78480 20 reactions
The Change-IT Multiple Mutation Site Directed Mutagenesis Kit is designed to create single or multiple oligonucleotide-directed sequence changes in plasmids.
�� Based on highly efficient exponential amplification using non-overlapping primers; eliminates primer-dimer formation�� Create single or multiple mutations in plasmids�� Create mutations in plasmids as large as 15 Kb�� Create insertions, base changes, or deletions�� Create deletions as large as 300 bp�� Encode multiple base changes in a single primer�� Three step process: amplification, Dpn I digestion, and transformation�� Single day method
Mutations are created using a variation of ligation-during-amplification(1) in which the plasmid is PCR-amplified with phosphorylated primers and the product ends are ligated together to convert the annealed DNA strands into closed circular DNA molecules (Fig. 1). These closed circular DNA molecules then become template molecules in subsequent rounds of the PCR amplification. The desired mutations (insertions, deletions, or substitutions) are created by incorporating them into single or multiple primer sequences. FideliTaq™ Polymerase is used to ensure faithful duplication of the plasmid sequence and successful long-distance PCR. Thermostable DNA ligase is utilized for efficient creation of circular plasmid DNA during each round of PCR amplification. For convenience, the enzymes are included in the kit as Change-IT Enzyme. Change-IT Enzyme is a blend of FideliTaq DNA Polymerase and a thermostable ligase in 50% glycerol. Together these enzymes generate exponentially amplified, circular, mutated plasmid DNA. To enhance selection of the mutated plasmid from the parental plasmid, the PCR amplification reaction is digested with Dpn I prior to transforma-tion into competent cells.
ConvenienceMinimal hands-on time is required – set up an amplification reaction, a restriction enzyme digest, and a transformation. Kit includes optimized reac-tion buffer, phosphorylated common primers to b-lactamase, and enzymes.
High fidelity and large plasmidsFaithful amplification of plasmids is ensured by FideliTaq Polymerase, included in the Change-IT enzyme blend, and an optimized Change-IT Buffer. Mutate plasmids as large as 15 kb.
High efficiencyExponential amplification of the mutated plasmid provides for high mutagenesis efficiency, even with multiple mutations.
Table 1. Change-IT mutagenesis efficiency
Number of Mutations % Mutated
Single 90
Double 90
Triple 80
The efficiency for the creation of one, two, or three mutations in a single template were determined for pUC19 (Table 1). The single mutation blocked expression of b-galactosidase (Fig. 2). The double mutation blocked expression of b-galactosidase and created a Xho I site. The triple mutation created an Nco I site, a Xho I site, and blocked expression of b-galactosidase. The single and double mutations were created with the greatest efficiencies. The triple mutation was present in 80% of the plasmid product, a lower rate than the double mutation as expected for this technique.
Kit components:10X Change-IT BufferChange-IT EnzymeDpn IAmp FWD Primer: PO4/CCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAmp REV Primer: PO4/GTCAGAAGTAAGTTGGCCGCAGTGTTATCACTCATGGMutagenic Control Primer:PO4/ATGACCATGATTACGCCATAGCTTGCATGCCTGCAGG0.5 ng/μl pUC19Nuclease-Free Water
Shipping and storage:Shipped on dry ice. Store at -20ºC.
Reference1. Chen, Z. and Ruffner, D.E. (1998) Nucl. Acids Res.
26(4), 1126-1127.
Change-IT Multiple Mutation Site Directed Mutagenesis Kit
Fig. 1. Change-IT Mutagenesis Method. Phosphorylated oligo-nucleotides are annealed to template plasmid. One oligonucleotide bears the desired sequence changes while the other anneals to a common sequence, such as the b-lactamase ORF. FideliTaq DNA Polymerase extends the oligonucleotides and DNA ligase seals the nascent DNA strands creating a replicated plasmid bearing the desired mutation. This process is repeated in a PCR amplification for 15 to 30 cycles, generating exponentially amplified, mutated, double-stranded plasmid as product for subsequent transformation.
P
Anneal and Extend
Ligate Cycle
Dpn I Digestion
Transformation
P
P
P
Multiple mutations
Fig. 2. Creation of multiple mutations. Single, double, or triple mutation reactions were performed. The single mutation inactivated LacZa, the double mutation inactivated LacZa and added a Xho I site, and the triple mutation inactivated LacZa and added both an Nco I site and a Xho I site. Images of plates demonstrate that the LacZa inactivating mutation was present in all conditions (i.e. white mutated colonies pre-dominate). Colony PCR amplifications were then analyzed by restriction digests. The product from the single mutation condition lacked both Nco I and Xho I sites, as expected (single band visible on agarose gel). Restriction analysis of the double mutation condition revealed two bands in all colonies screened, indicating all carried the Xho I site insertion. Restriction analysis of the triple mutation condition revealed that 9 of 10 colonies incorporated all three site-directed mutations.
127For bulk or alternate pack sizes, email us at [email protected].
Molecular Biology Products & Kits | Labeling
78430 30 reactions
The Sequenase Random Primer Labeling Kit offers a rapid and convenient method to generate DNA probes with high specific activity, >1.5 x 109 dpm/μg. The kit features Sequenase Version 2.0 DNA Polym-erase and random nonamers (9-mers) in a protocol that has been optimized to produce radiolabeled probes in 5 minutes. The labeled probes may be used for screening of gene libraries(1), Southern and North-ern blots(2,3), as well as in situ hybridizations.
�� Rapid, 5-minute reaction�� DNA probes labeled to high specific activity, >1.5 x 109 dpm/μg�� Easy-to-use 10X Nucleotide Mixes�� Flexible labeling with either radiolabeled dATP or dCTP�� Optimized labeling conditions using Sequenase Version 2.0 DNA Polymerase and random nonamers
Random primer labeling methodRandom primed labeling of DNA was originally developed by Feinberg and Vogelstein and used Klenow DNA Polymerase as the enzyme for polym-erization(4,5). The method relies on annealing random DNA primers along the length of a target DNA so that the primed DNA can be used as a substrate for the polymerase. The DNA is radiolabeled by substituting one of the nucleotides of the reaction with a radioactive one ([32P], [33P], or [35S]), and the resulting radiolabeled DNA can be used as probes in various hybridization techniques such as Southern and Northern blots and in situ hybridizations. This original protocol has been enhanced by labeling probes with Sequenase DNA Polymerase instead of Klenow(6). Sequenase DNA Polymerase is the enzyme of choice because of its superior activity in primer extension reactions and its lack of exonuclease activ-ity that tends to degrade radiolabeled probes. Also, the kit includes random nonamers (9-mers) instead of hexamers (6-mers) for the labeling reaction to provide more efficient priming of the target DNA(7).
Applications:�� Southern blots�� Northern blots�� Solution hybridization�� In situ hybridization
High specific activity probesThe Sequenase Random Primer Labeling Kit suc-cessfully labels varying amounts of DNA up to approximately 2 μg, but 25 ng is recommended for maximal probe specific activity. Figure 1 shows a time course of the labeling reaction using 25 ng of Lambda DNA that was monitored by the incorpora-tion of [33P]-dATP. The standard protocol routinely gives a specific activity of >1.5 x 109 dpm/µg within five minutes, corresponding to an incorporation of up to 70% of the radioactive nucleotide.
Consistent probe sizeThe Sequenase Random Primer Labeling Kit gener-ates a consistent range of probe sizes on a variety of DNA templates, including lambda DNA, pUC18 plasmid DNA, as well as DNA fragments gener-ated by PCR (Fig. 2). The labeling protocol routinely produces DNA probes ranging in size from 100-850 nucleotides in length.
Easy to useThe Sequenase Random Primer Labeling Kit provides two different 10X Nucleotide Mixes (-dATP and -dCTP) for flexible labeling. Using these buffers, DNA probes can be generated equally well using either radioactively labeled dATP or dCTP.
Kit components: 10X Primer Mix10X Nucleotide Mix, minus dATP10X Nucleotide Mix, minus dCTPSequenase Version 2.0 DNA PolymeraseWater, Nuclease-FreeStop Buffer (0.5 M EDTA)Control DNA (lambda DNA)
Shipping and storage:Shipped on dry ice. Store at -20ºC.
References: 1. Grunstein, M. and Hogness, D. S. (1975) Proc.
Natl. Acad. Sci. USA 72, 3961-3965.2. Southern, E. M. (1975) J. Mol. Biol. 98, 503-517.3. Smith, G. E. and Summers, M. D. (1980) Anal.
Biochem. 109, 123-129.4. Feinberg, A. P. and Vogelstein, B. (1983) Anal.
Biochem. 132, 6-13.5. Feinberg, A. P. and Vogelstein, B. (1984) Anal.
Biochem. 137, 266-267.6. U.S. Pat. No. 4,795,699.7. Stangegaard, M., Dufva, I. H., and Dufva, M.
(2006) BioTechniques 40, 649-657.
Sequenase Random Primer Labeling Kit
Fig. 2. Size comparison of DNA probes generated from lambda DNA (Lane 1), linearized pUC18 (Lane 2), and a PCR fragment, 1.6 kb (Lane 3).
Fig. 1. Specific activity of DNA probes generated from lamb-da DNA through time.
Minutes
Spec
ific
Activ
ity(x
10
9dp
m/
g)
1510500.0
0.5
1.0
1.5
2.0
2.5
850 —
500 —400 —300 —200 —
100 —
DNA probesMW (nt)
1 2 3
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128 888-362-2447 | 216-765-5000 | usb.affymetrix.com
DNA, Calf ThymusULTRAPURE
Source: Calf ThymusDescription: Highly polymerized DNA; Characterized
as a high molecular weight DNA prepared by ethanol precipitation. This DNA is an excellent substrate for DNase.
Product specifications:Assay: Hyperchromicity ≥ 27%A260/A280: ≥ 1.8
Storage: 4°C, desiccatedCAS #9007-49-2
14375 1 gm 5 gm
DNA, E. coli
Source: E. coliDescription: E. coli genomic DNA is purified from
E. coli type B cells, ATCC 11303 strain. It has a single chromosome and the genome is 4,600,000 bp long. This genomic DNA is fragmented to some degree during purification yet it is charac-terized as a high molecular weight DNA.
Product specifications:Form: Dried powderHyperchromicity: ≥ 27%A260/A280: ≥ 1.8
Storage: 4°CCAS #9007-49-2
14380 10 mg
DNA, Fish Sperm
Source: Fish SpermProduct specifications:Form: Off-white to yellowish/brownish powderMoisture: ≤ 7%Nitrogen: ≥ 14.5%Phosphorous: ≥ 9.0%Protein: NegativeCAS #100403-24-5
14405 100 gm 1 kg
DNA, Fish Sperm, Activated
Form: Solution in 10 mM Tris-HCI, pH 7.5, 1 mM EDTA.
Activity Analysis: This DNA gives the optimum activ-ity in standard assay for Sequenase Version 2.0 DNA Polymerase.
Storage: -20°C
70076 500 µg 2 mg
DNA, Fish Sperm, Sodium Salt
Source: Fish SpermProduct specifications:Form: Off-white to light tan powderPhosphorous: ≤ 9.0%Protein: ≤ 5.0%Loss on Drying: ≤ 18.0%CAS #68938-01-2
14377 100 gm 1 kg
DNA-Cellulose, Double-Stranded(dsDNA-Cellulose)
Source: Calf ThymusDescription: Calf Thymus DNA (PN 14375) and ash
free, acid washed, chromatographic grade cellu-lose and crosslinked by ultraviolet irradiation.
Product specifications:Form: Dried powder (may contain traces of NaCl,
Tris-HCl and EDTA).Use: Column chromatography media for the iso-
lation of DNA binding proteins from various sources.
Note: Store as dry powder after washing with H2O and ethanol.
Note: One gram of DNA Cellulose will swell to 3-4 ml gel when fully hydrated.
CAS #9004-34-6
14394 5 gm 10 gm
Molecular Biology Products & Kits | DNA
We have been providing custom enzymes and bulk biochemicals for more than 40 years, helping academic institutions, diagnostic, biotech, and pharmaceutical companies succeed. We combine the resources of a large organization with the personalized service of a dedicated team. Turn to us for:
� Custom reagent manufacturing, dispensing, labeling, and kitting
� 3,000 catalog enzymes and biochemicals that can be tailored to meet your specific needs
� An ISO 13485 certified quality system well-suited to meet diagnostic requirements
� In-house experts in custom and OEM reagents to guide you through the manufacturing process
129For bulk or alternate pack sizes, email us at [email protected].
DNA-Cellulose, Single-Stranded(ssDNA-Cellulose)
Source: Calf ThymusDescription: Calf Thymus DNA (PN 14375) and ash
free, acid washed, chromatographic grade cellu-lose and crosslinked by ultraviolet irradiation.
Product specifications:Form: Dried powder (may contain traces of NaCl,
Tris-HCl and EDTA). One gram of powder swells to approx. 3 ml of final hydrated media.
Note: Store as dry powder after washing with H2O and ethanol.
Use column chromatography media for the isolation of DNA binding proteins from various sources.
Note: One gram of DNA Cellulose will swell to 3-4 ml gel when fully hydrated.
CAS #9004-34-6
14395 5 gm 10 gm
M13mp18, Single-Stranded, 0.2 µg/µl
Source: E. coli strain JM101 infected with bacterio-phage M13mp18
Storage Buffer: Supplied in 10 mM Tris-HCl (pH 8.0), 1 mM EDTA.
Concentration: 0.2 µg/µl
70704 50 µl 50 µg
M13mp18 DNA, Single-Stranded, 1.0 µg/µl
Source: E. coli strain JM101 infected with bacterio-phage M13mp18
Storage Buffer: Supplied in 10 mM Tris-HCl (pH 7.5), 1 mM EDTA.
Concentration: 1.0 µg/µl
71706 50 µg 200 µg
Oligo(dT)12-18 Primer, MW ≅ 4500
MB GradeForm: 25 µM ± 2.5 in dH2O.2.5 nmol per tube.Storage: -20°C.
77405 100 µl (2.5 nmol)
Oligo p(dT)12-18 Sodium Salt
Form: Lyophilized powderDescription: This oligodeoxythymidylate mixture is
synthesized using O-cyanoethyl phosphoramidite chemistry. After the removal of the protecting groups, the product is converted to the sodium salt and the excess salt removed by gel filtration.
Unit Definition: One A260 unit is that quantity of material which, when dissolved in 1.0 ml of water and read in a spectrophotometer at 260 nm in a cuvette with a 1.0 cm light path, gives an absor-bance of 1.0. One A260 unit is approximately equal to 30-35 µg of DNA.
Storage: -20°C
19817 5 units 25 units 100 units
pUC19 DNA, 0.2 µg/µl
Storage Buffer: Supplied in 10 mM Tris-HCl (pH 7.5), 1 mM EDTA.
Concentration: 0.2 µg/µlFunctional Test: Tested in DNA sequencing
76632 10 µg 50 µg 200 µg
pUC19 DNA, 1.0 µg/µl
Storage Buffer: Supplied in 10 mM Tris-HCl (pH 7.5), 1 mM EDTA.
Concentration: 1.0 µg/µlFunctional Test: Tested in DNA sequencing
76634 50 µg 200 µg
Custom sizes, concentrations and pack sizes available upon request
Molecular Biology Products & Kits | DNA
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Molecular Biology Products & Kits | DNA Markers
76710 250 µl
6X DNA Loading Buffer (OXG)76715 1 ml 5 ml
Description:PCR Markers are a mixture of 8 DNA bands ranging in size from 50 to 2000 bp in ready-to-load form supplied in gel loading buffer. The recommended amount for loading (5 µl) produces bands of high resolution for easy size estimation of PCR products.
6X DNA Loading Buffer (OXG) with orange G and xylene cyanol FF is included for convenience to pre-pare PCR samples for agarose gel electrophoresis. The tracking dyes do not interfere with UV illumina-tion of ethidium bromide stained gels.
Fragments: 2000 bp 500 bp 1500 bp 300 bp 1000 bp 150 bp 750 bp 50 bp
To use: Add 5 µl PCR Marker per gel lane.
6X DNA Loading Buffer (OXG) (PN 76715, 1 ml included):6X Loading Buffer consists of orange G, xylene cyanol FF, and glycerol. Shipped ambient.
Functional test:Meets or exceeds criteria for agarose gel electro-phoresis.
Shipping and storage:Shipped on ice. Store at 4°C.
PCR Markers, 50 - 2000 bp
– 2000 – 1500
– 1000 – 750
– 500
– 300
– 150
– 50
bp
1.5% TAE agarose gel
76712 500 µl
6X DNA Loading Buffer (OXG)76715 1 ml 5 ml
Description:The 100 bp DNA Ladder is a set of 10 easy-to-re-member band sizes for use in qualitative agarose gel analysis. The ladder is supplied in ready-to-load form in gel loading buffer. Simply add the recommended amount (5 µl) for bright band intensities between 100 and 1000 base pairs.
6X DNA Loading Buffer (OXG) with orange G and xylene cyanol FF is included for convenience to prepare DNA samples for agarose gel electrophore-sis. The tracking dyes do not interfere with UV illumination of ethidium bromide stained gels.
Fragments: 1000 bp 500 bp 900 bp 400 bp 800 bp 300 bp 700 bp 200 bp 600 bp 100 bp
To use: Add 5 µl of ladder mix per gel lane.
6X DNA Loading Buffer (OXG) (PN 76715, 1 ml included):6X Loading Buffer consists of orange G, xylene cyanol FF, and glycerol. Shipped ambient.
Functional test:Meets or exceeds criteria for agarose gel electro-phoresis.
Shipping and storage:Shipped on ice. Store at 4°C.
DNA Ladder, 100 bp
2.0% TAE agarose gel
– 1000 – 900 – 800 – 700 – 600 – 500
– 400
– 300
– 200
– 100
bp
131For bulk or alternate pack sizes, email us at [email protected].
Molecular Biology Products & Kits | DNA Markers
76714 500 µl
6X DNA Loading Buffer (BXF)76720 1 ml 5 ml
Description:The 1 kb Plus DNA Ladder is a set of 14 easy-to-remember band sizes for use in qualitative agarose gel analysis ranging in size from 12 kb down to 100 bp. Two high intensity reference bands of 4 kb and 500 bp are included. The ladder is supplied in ready-to-load form in gel loading buffer. Simply add the recommended amount (5 µl) for highly resolved bands.
6X DNA Loading Buffer (BXF) with bromophenol blue and xylene cyanol FF is included for conve-nience to prepare DNA samples for agarose gel electrophoresis.
Fragments: 12 kb 1.5 kb 10 kb 1 kb 8 kb 500 bp 6 kb 400 bp 4 kb 300 bp 3 kb 200 bp 2 kb 100 bp
To use: Add 5 µl ladder mix per gel lane.
6X DNA Loading Buffer (BXF) (PN 76720, 1 ml included):6X Loading Buffer consists of bromophenol blue, xylene cyanol FF, and Ficoll®.
Functional test:Meets or exceeds criteria for agarose gel electro-phoresis.
Shipping and storage:Shipped on ice. Store at 4°C.
DNA Ladder, 1 kb Plus
1.0% TAE agarose gel
– 12,000 – 10,000 – 8000 – 6000
– 4000 – 3000
– 2000
– 1500
– 1000
– 500 – 400 – 300 – 200 – 100
bp
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Molecular Biology Products & Kits | Oligonucleotide Marker
76410 100 µl
Description:The Low Molecular Weight Marker is a set of 14 bands between 10 - 100 nucleotides, designed for size determination of single-stranded oligonucleotide fragments.
The marker is suitable for use in either polyacryla-mide or agarose gel electrophoresis. The recom-mended concentration for agarose is 2.5% and for polyacrylamide is 12 - 15%. This marker should be diluted with TE Buffer and the supplied Gel Loading Buffer to the desired loading concentration.
The Low Molecular Weight Marker can be used to generate a 5’-end radiolabeled marker for size comparison of small, single-stranded oligonucleotide fragments such as miRNA, siRNA, tRNA, snRNA, snoRNA, or ssDNA. The Low Molecular Weight Marker is also useful when the molecule of interest is less than 100 nucleotides in length. This marker is not designed for precise quantification of nucleic acid mass and should only be used as a size marker.
2X Formamide Loading Buffer and 6X Glycerol Load-ing Buffer are included for convenience to prepare both the marker and samples for gel electrophoresis.
Fragment sizes: 100 nt 40 nt 90 nt 35 nt 80 nt 30 nt 70 nt 25 nt 60 nt 20 nt 50 nt 15 nt 45 nt 10 nt
Functional test: Meets or exceeds criteria for 5'-end labeling with isotope and polyacrylamide gel electrophoresis.
Components:Low Molecular Weight Marker, 100 µl (50 ng/µl each
fragment)2X Formamide Loading Buffer6X Glycerol Loading Buffer
2X Formamide Loading Buffer (1 ml included):Consists of bromophenol blue, xylene cyanol FF, EDTA and formamide.
6X Glycerol Loading Buffer (1 ml included): Consists of orange G, xylene cyanol FF and glycerol.
Shipping and storage:Shipped on ice. Store at 4°C or at -20°C for long term storage.
To use:1. Mix 2 µl (100 ng each fragment) of Marker with
2 µl of 2X Formamide Loading Buffer, heat at 95°C for 3 minutes, quick cool on ice, and load the entire volume per lane on polyacrylamide gel.
2. Mix 5 µl (250 ng each fragment) of Marker with 1 µl of 6X Glycerol Loading Buffer and load the entire volume per lane on agarose gel.
3. Visualize with dyes for detecting single-stranded oligonucleotide fragments.
4. For 5'-end labeling reaction, use 1 µl (50 ng each fragment) of Marker with OptiKinase™ and [γ-32P]-ATP or [γ-33P]-ATP.
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Low Molecular Weight Marker, 10 – 100 nt
Fig. 1. Low Molecular Weight Marker on 12-15% UREA-polyacrylamide gel. A. 50 ng of SYBR® Gold stained Marker. B. 0.5 ng of 5' end labeled Marker with [γ-33P]-ATP and OptiKinase. C. 0.05 ng of 5' end labeled Marker with [γ-32P]-ATP and OptiKinase.
100908070
60
5045
40
35
30
25
20
15
10
100908070
60
5045
40
35
30
25
20
15
10
10090807060
5045
40
35
30
25
20
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10
A B Cnt nt nt
133For bulk or alternate pack sizes, email us at [email protected].
Molecular Biology Products & Kits | Protein Analysis
Human Angiogenesis Antibody ArrayMA6310 kitMouse Angiogenesis Antibody ArrayMA6320 kit
�� Simultaneously profile expression of multiple proteins associated with angiogenesis�� Analyze the effects of different stimuli on the induction or inhibition of angiogenesis
With Angiogenesis Antibody Arrays, you can detect multiple angiogenesis-specific cytokines in a single hybridization experiment. These arrays are extremely sensitive, detecting proteins at concentrations in the pg/ml range.
Angiogenesis, the generation of blood vessels from endothelial cells, is normally controlled by pro-angio-genic “activator” molecules and anti-angiogenic “inhibitor” molecules. In some cancers, however, the careful balance of these molecules is upset, as tumor cells promote vascularization to obtain nutrients and oxygen necessary for growth. Tumor growth is sustained by abnormal levels of these angiogenesis-related proteins; the affected proteins vary from one type of cancer to another. To gain a clear picture of the angiogenesis-stimulating pathways used by cancer cells, it is necessary to investigate the levels of these cytokines in tumors. With the Angiogenesis Antibody Arrays, you can simultaneously profile 19 angiogen-esis activators and inhibitors at the protein level.
We currently offer Angiogenesis Antibody Arrays for both human and mouse. These arrays can be used with a variety of biological sources, including cell extracts, tissue lysates, conditioned media, patient sera, and plasma.
Kit components:Four array membranesBiotinylated angiogenesis antibodiesBlocking, hybridization, and wash solutionsHRP detection reagents
Angiogenesis Antibody Array content Here’s how it works
Angiogenesis Antibody Arrays
Human Angiogenesis Array
Angiogenin
G-CSF
HGF
Leptin
VEGF
IL-1a
IL-1b
IL-6
IL-8
PIGF
FGFa
FGFb
TNFa
TGFa
IFNγ
IL-12
IP-10
TIMP-1
TIMP-2
Mouse Angiogenesis Array
EGF
FGFa
FGFb
G-CSF
IL-1a
IL-1b
IL-4
IL-6
Leptin
VEGF
TNFa
TGFa
TGFb
IFNγ
IL-12
IP-10
TIMP-1
TIMP-2
The Angiogenesis Antibody Array profiles expression of angiogenesis-specific cytokines. Tissue extracts from normal (Panel A) and cancerous (Panel B) human colon were incubated with the Angiogenesis Antibody Array membrane. Chemiluminescent images were collected using a FluorChem imager (Alpha Innotech), revealing increases in the expression of certain cytokines in the can-cerous tissue extract (boxes).
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Looking to profile cytokines from a larger number of samples? Call our Technical Support team and ask about the Procarta Cytokine Plex Profiling Kits. Both human and mouse panels are available.
134 888-362-2447 | 216-765-5000 | usb.affymetrix.com
AFP ELISA Kit BC1009 kitCA125 ELISA Kit BC1013 kitCA15-3 ELISA Kit BC1015 kitCA19-9 ELISA Kit BC1017 kit
Quickly quantify cancer antigen concentra-tions in human serumCancer Antigen ELISA Kits allow rapid screening of up to 96 samples at once for quantitative determina-tion of tumor marker concentration. This ELISA technology allows you to quickly and efficiently screen multiple human serum samples for various antigens shown to be present at elevated levels in different cancers. The four assays we offer are for alpha-fetoprotein (AFP) and cancer antigens CA125, CA15-3, and CA19-9.
Note: The Cancer Antigen ELISA Kits are intended for research use only.
Kit components:One 96-well microtiter plate coated with anti-cancer
antigenProtein standardsComplete reaction and detection reagents
AFPElevation of serum AFP to abnormally high levels occurs in several malignant diseases (including nonseminomatous testicular cancer and primary hepatocellular carcinoma) and some benign ones (including hepatitis and cirrhosis).
CA125Cancer Antigen 125 (CA125) is a surface antigen associated with epithelial ovarian cancer, and to date CA125 is the most sensitive marker for residual epi-thelial ovarian cancer. CA125 may also be elevated in patients with lung, cervical, fallopian tube, and uterine cancers, as well as endometriosis.
CA15-3Cancer antigen 15-3 (CA15-3) is useful for monitor-ing breast cancer patients post-operatively for recurrence, particularly with metastatic disease. CA15-3 levels are also increased in colon, lung, and hepatic tumors.
CA19-9Serum CA19-9 level is frequently elevated in subjects with certain gastrointestinal malignancies, such as pancreatic, colorectal, gastric, and hepatic carcino-mas. A consistently rising serum CA19-9 value may be associated with progressive malignant disease and poor therapeutic response. A declining CA19-9 value may be indicative of a favorable prognosis and good response to treatment.
Cancer Antigen ELISA Kits
AFP
0.0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
0 50 100 150 200 250 300
Concentration ng/mlA
450
A45
0
CA125
0.0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
0 100 200 300 400
Units/ml
A45
0
CA15-3
Units/ml
0.0
0.5
1.0
1.5
2.0
2.5
3.0
0 50 100 150 200 250
CA19-9
A45
00.0
0.5
1.0
1.5
2.0
2.5
3.0
0 100 200 300 400 500 600
Units/ml
Human Cytokine Antibody Array 1.0MA6110 2 membranesHuman Cytokine Antibody Array 1.0MA6120 4 membranesHuman Cytokine Antibody Array 1.0MA6130 6 membranesHuman Cytokine Antibody Array 1.0MA6140 8 membranesHuman Cytokine Antibody Array 3.0MA6150 4 membranesHuman Cytokine Antibody Array 3.0MA6160 8 membranesMouse Cytokine Antibody Array 1.0MA6410 4 membranesMouse Cytokine Antibody Array 1.0MA6412 8 membranes
�� Simultaneously profile the expression of multiple cytokine proteins.�� Analyze regulation of cytokine expression with different cellular stimuli.
With the Cytokine Antibody Arrays, you can simul-taneously detect multiple cytokines from a variety of sources, including serum, conditioned media, plasma, and cell/tissue lysates. While cDNA arrays give information about levels of gene expression, the Cytokine Antibody Arrays detect the levels of active cytokine protein.
The Cytokine Antibody Arrays have several advan-tages over traditional methods such as ELISA. The sensitive high-throughput assay is a key benefit—these arrays can detect soluble cytokine proteins at concentrations in the pg/ml range. There is also less variability with these arrays compared to ELISAs; variability between the same spots on two of the same type of membrane is typically less than 10%, while the variation between ELISA plates is about 20%. Most importantly, the Cytokine Antibody Ar-rays facilitate the simultaneous detection of multiple cytokines in one assay.
We currently offer three of these arrays—two for human and one for mouse. The Human Cytokine Antibody Array comes in two formats: Version I.0 includes 18 cytokines and Version 3.0 includes 36 cytokines. The Mouse Cytokine Antibody Array 1.0 includes 18 cytokines.
The Cytokine Antibody Arrays can be purchased with different numbers of membranes.
Kit components:Array membranesBiotinylated anti-cytokine antibodiesStreptavidin-HRP conjugateBlocking and wash solutionsHRP detection reagents
Cytokine Antibody Arrays
Molecular Biology Products & Kits | Protein Analysis
135For bulk or alternate pack sizes, email us at [email protected].
The Human Cytokine Antibody Array 3.0 detects differences among active cytokines from A431 cells exposed to different conditions. Membranes were incubated with conditioned media from untreated A431 cells (Panel A) and A431 cells treated with EGF (Panel B) or PMA (Panel C). Images were collected using a FluorChem imager (Alpha Innotech). Differences in protein expression were observed among the untreated and treated samples, as indicated in boxes.
B. Unstimulated A431 and EGF C. Stimulated A431 and PMAA. Unstimulated A431
Human Cytokine Array 1.0
EotaxinIL-1aIL-10GM-CSFIL-1bIL-12IP-10IL-3IL-17TNFaIL-4MIP1aRantesIL-6MIP1bLeptinIL-8MIP-5
Mouse Cytokine Array 1.0
G-CSFM-CSFGM-CSFMIGMIP-1aIFNγ
TNFaIP-10RantesVEGFIL-1aIL-2IL-4IL-5IL-6IL-10IL-12IL-13
Human Cytokine Array 3.0
Apol/FasCTLAEotaxinGM-CSFEGFIP-10LeptinMIP1aMIP1bMIP4MIP-5MMP3RantesTGFbIFNγ
TNFaTNFRITNFRIIICAM-1VCAM-1VEGFIL-1aIL-1bIL-1RaIL-2IL-3IL-4IL-5IL-6IL-6RIL-7IL-8IL-10IL-12 (p40)IL-15IL-17
Here’s how it works
Cytokine Antibody Arrays continued...
Cytokine Antibody Content
Molecular Biology Products & Kits | Protein Analysis
136 888-362-2447 | 216-765-5000 | usb.affymetrix.com
76740 500 µl
Description:The 10 - 225 kDa Protein Markers are recombinant proteins designed for convenient and accurate sizing of protein samples. Unlike conventional protein markers, the recombinant marker proteins do not contain oligosaccharides which often cause poor resolution and inaccurate size estimation of protein bands.
An added benefit is the ability to estimate the con-centration of protein samples using the known mass for each protein band. Each vial (500 µl) contains 500 µg of protein - 50 µg per band except for 100 µg of the 50 kDa high intensity reference band.
The markers are in ready-to-load form in gel loading buffer. Simply add 5 µl of the marker to a standard SDS-polyacrylamide gel for visualization with Coomassie blue.
As an added convenience, 2X SDS Sample Buffer is included for preparing protein samples for gel electrophoresis.
Protein bands (MW): 225 kDa 35 kDa 150 kDa 25 kDa 100 kDa 15 kDa 75 kDa 10 kDa 50 kDa
The 50 kDa band is a high intensity reference band.
Storage buffer:1 µg/µl total marker protein in 125 mM Tris-HCl, pH 6.8, 2% SDS, 10% glycerol, 200 mM b-ME, 0.007% bromophenol blue.
To use: Add 5 µl of marker per gel lane on standard SDS-polyacrylamide gels for staining with Coomassie blue. For silver staining or S-Taq analysis, use 5 µl of a 1:25 dilution in 1X Sample Buffer.
2X SDS Sample Buffer (1 ml included)250 mM Tris-HCl, pH 6.8, 6% SDS, 300 mM DTT, 30% glycerol, and 0.02% bromophenol blue.
Functional test:Meets or exceeds criteria for SDS-PAGE.
Shipping and storage:Shipped on ice. Store at 4°C.
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RapidGel, 40% Liquid Acrylamide Stock Solution75848 500 ml
Sodium Dodecyl Sulfate, 10% Solution77504 100 ml 500 ml 1 L
TBS with Tween (TBST), 20X Solution77500 1 L 5 L
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Tris-Glycine (TG) Buffer, 10X Solution75894 1 L 5 L
Tris-Glycine-SDS (TGS) Buffer, 10X Solution75895 1 L 5 L
Protein Markers, 10 - 225 kDa
Stained with Coomassie Blue 8-16% Tris-Glycine gel.
kDa – 225
– 150
– 100
– 75
– 50*
– 35
– 25
– 15 – 10
Molecular Biology Products & Kits | Protein Markers
137For bulk or alternate pack sizes, email us at [email protected].
76230Y 100 mg76230Z 500 mg76230 1 gm (2 x 500 mg)Customs by request
Source:Tritirachium album
Description:Proteinase K is a highly active and stable endopep-tidase purified from the fungus T. album. It cleaves peptide bonds mainly following the carboxyl group of N-substituted hydrophobic aliphatic and aromatic amino acids(1) and is classified as a serine protease. Proteinase K is useful in purifying high molecular weight nucleic acids from cells and tissues (i.e. genomic DNA isolation from mouse tails) and in revealing protein structure and function.
Applications:�� Used in the isolation of native, high molecular weight DNA and RNA.�� Used as a general protease to inactivate DNases, RNases, and to degrade proteins.�� Specifically modifies cell surface proteins and glycoproteins for analysis of membrane structures for protein localization(5,6).�� Produces characteristic protein fragments used in enzyme/protein structure and function studies(7).
Form:Lyophilized powder
Properties:Molecular Weight: 28.9 kDaOptimum pH: 4.0-12.5(1)
Optimum Temperature: 65°C(1)
Working Concentration: 0.05 - 1 mg/mlActivators: Stimulated by 0.2 - 1% SDS or 1 - 4 M
urea(8) and 1 - 5 mM Ca2+
Inactivated by PMSF and diisopropyl phosphoro-fluoridate (DPF). Partially inactivated by EGTA. Not inactivated by EDTA, thiol reagents, or trypsin/chymotrypsin inhibitors(1)
Note: Proteinase K has two binding sites for Ca2+ which act as a stabilizing factor of the enzyme. When Ca2+ is removed from the enzyme approxi-mately 80% of the catalytic activity is lost because of the structural change(2). However, residual Proteinase K activity is usually sufficient to degrade proteins commonly present during DNA/RNA isolation.
Purity:Tested for contaminating ribonucleases and deoxy-ribonucleases.
Unit definition:One unit is the amount of enzyme that liberates folin positive amino acids and peptides corresponding to
1 µmol tyrosine in 1 minute at 37°C using hemoglo-bin as substrate.
Concentration:20 - 50 units/mg
Shipping and storage:Shipped on cold pack. Store dry at 4°C.
References:1. Ebeling, W., Hennrich, N., Klockow, M., Metz, H.,
Orth, H. D. and Lang, H. (1974) Eur. J. Biochem. 47, 91-97.
2. Gross-Bellard, M., Oudet, P. and Chambon, P. (1973) Eur. J. Biochem. 36, 32-38.
3. Hansen, J. N. (1974) Prep. Biochem. 4, 473-488.4. Kasche, V., Zollner, R., Amneus, H. and Naslund, L.
(1981) Prep. Biochem. 11, 233-250.5. Roelcke, D. and Ulenbruck, G. (1969) Z. Med.
Mikrobiot. und Immunol. 155, 156-170.6. Brdiczka, D. and Krebs, W. (1973) Biochem.
Biophys. Acta 297, 203-212.7. Williamson, J., Greene, J., Cherif, S. and Milner-
White, E. J. (1977) Biochem. J. 167, 731-737.8. Hilz, H., Wiegers, U. and Adamietz, P. (1975) Eur. J.
Biochem. 56, 103-108.9. Crowe, J. S., Cooper, H., Smith, M., Sims, M.,
Parker, D. and Gewert, D. (1991) Nucl. Acids. Res. 19, 184.
Proteinase K
76225 100 mg
Source:Tritirachium album
Description:Proteinase K is a highly active and stable endopep-tidase purified from the fungus T. album. It cleaves peptide bonds mainly following the carboxyl group of N-substituted hydrophobic aliphatic and aromatic amino acids(1) and is classified as a serine protease. Proteinase K is useful in purifying high molecular weight nucleic acids from cells and tissues and in revealing protein structure and function.
Applications:�� Isolation of DNA and RNA to inactivate DNases, RNases and to degrade other proteins during cell digestion.�� Useful in the specific modification of cell surface proteins and glycoproteins for analysis of membrane structures(4,5).�� Useful in obtaining characteristic protein fragments which help reveal the structure and function of enzymes and other proteins(6).
Properties:Molecular Weight: 28,900 DapH Range: 4.0 -12.5(1) with optimum activity at
pH 8.0Temperature Range: 37- 60°C(1)
Working Concentration: 0.05 - 1 mg/ml
Stabilizer: Ca2+; Activity decreases if Ca2+ is removed from the solution(2)
Activity: Active in the presence of 1% SDS, 4 M Urea(3), 1% Triton X-100(3), 5% Tween-20, or 3 M Guanidine HCl.
Inactivated by PMSF and diisopropyl phosphoro-fluoridate (DPF). Partially inactivated by EGTA. Not inactivated by EDTA, thiol reagents, or trypsin/chymotrypsin inhibitors(1)
Inactivation: Inactivate by heating at 95°C for 10 minutes, PMSF or DPF.
Purity:Tested for contaminating exonucleases, endonucle-ases, and ribonucleases.
Storage buffer:20 mM Tris-HCl (pH 7.4), 1.0 mM CaCl2, and 50% glycerol.
Unit definition:One unit is the amount of enzyme that liberates folin positive amino acids and peptides corresponding to 1 µmol tyrosine in 1 minute at 37°C using hemoglo-bin as a substrate.
Activity: ≥ 600 units/ml
Specific activity: ≥ 30 units/mg
Protein concentration: 20 mg/ml
Recommended reaction buffer: 50 mM Tris-HCl (pH 7.4) and 1.0 mM CaCl2 and 50% glycerol.
Note: Proteinase K has two binding sites for Ca2+ which act as a stabilizing factor of the enzyme. When Ca2+ is removed from the enzyme approxi-mately 80% of the catalytic activity is lost because of the structural change(2). However, residual Proteinase K activity is usually sufficient to degrade proteins commonly present during DNA/RNA isolation.
Shipping and storage:Shipped on dry ice. Store at -20°C.
References:1. Ebeling, W., Hennrich, N., Klockow, M., Metz, H.,
Orth, H. D. and Lang, H. (1974) Eur. J. Biochem. 47, 91-97.
2. Sambrook, J., Fritsch, E. F. and Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual, 2nd Edition, Cold Spring Harbor, New York.
3. Hilz, H., Wiegers, U. and Adamietz, P. (1975) Eur. J. Biochem. 56, 103-108.
4. Roelcke, D. and Ulenbruck, G. (1969) Z. Med. Mikrobiot. und Immunol. 155, 156-170.
5. Brdiczka, D. and Krebs, W. (1973) Biochem. Biophys. Acta 297, 203-212.
6. Williamson, J., Greene, J., Cherif, S. and Milner-White, E. J. (1977) Biochem. J. 167, 731-737.
Proteinase K Solution
Molecular Biology Products & Kits | Proteinases
PurificationPlasmid Purification
Midi to Giga Preps
Spin Columns
Endotoxin-Free Preps
Medium Throughput
High Throughput
Yeast
BAC
DNA Cleanup
Gel Extraction
Genomic DNA Isolation
PCR Purification
RNA Purification
Sequencing Dye Cleanup
Histidine-Tagged Protein
Purification Kits & Resins
GST-Tagged Protein Purification
139For bulk or alternate pack sizes, email us at [email protected].
Purification | Table of Contents
Plasmid Purification – Midi to Giga PrepsPrepEase® Midi Plasmid Kit 140PrepEase Maxi Plasmid Kit 140PrepEase Giga Plasmid Kit 140
Plasmid Purification – Spin ColumnsPrepEase Quick MiniSpin Plasmid Kit 141PrepEase MiniSpin Plasmid Kit 141
Plasmid Purification – Endotoxin-Free PrepsPrepEase Endotoxin-Free Maxi Plasmid Kit 142
Plasmid Purification – Medium ThroughputPrepEase Plasmid 8-well Strip Kit 143PrepEase 8-well Strip Starter Kit for Vacuum 143PrepEase Vacuum Manifold 143
Plasmid Purification – High ThroughputPrepEase Plasmid 96-well Plate Kit 144PrepEase Plasmid 96-well Plate Core Kit 144
Plasmid Purification – YeastPrepEase Yeast Plasmid Isolation Kit 145
Plasmid Purification – BACPrepEase BAC Purification Kit 145
DNA CleanupPrepEase DNA Cleanup Kits 146
Gel ExtractionPrepEase Gel Extraction Kits 146
Genomic DNA IsolationPrepEase Genomic DNA Isolation Kits 147
PCR PurificationPrepEase PCR Purification 96-Well Plate Kits
(Ultrafiltration) 148ExoSAP-IT® PCR Product Cleanup 148HT ExoSAP-IT High-Throughput PCR Product
Cleanup 149
RNA PurificationPrepEase RNA Spin Kits 150PrepEase mRNA MiniSpin Kit 150PrepEase RNA/Protein Spin Kit 151
Sequencing Dye CleanupPrepEase Sequencing Dye Cleanup Kits 152
Histidine-Tagged Protein Purification KitsPrepEase Histidine-Tagged Mini Kit – High
Specificity 153PrepEase Histidine-Tagged Midi Kit – High
Specificity 153PrepEase Histidine-Tagged Maxi Kit – High
Specificity 153PrepEase Histidine-Tagged Mini Kit – High
Yield 154PrepEase Histidine-Tagged Midi Kit – High
Yield 154PrepEase Histidine-Tagged Maxi Kit – High
Yield 154
Histidine-Tagged Protein Purification ResinsPrepEase Histidine-Tagged High Specificity
Purification Resin 155PrepEase Histidine-Tagged High Yield
Purification Resin 155
GST-Tagged Protein PurificationPrepEase Protein Purification Glutathione
Agarose 4B 155
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Purification | Plasmid Purification – Midi to Giga Preps
PrepEase Midi Plasmid KitBinding capacity up to 100 µgCulture volume up to 30 ml78706 100 preps
PrepEase Maxi Plasmid KitBinding capacity up to 500 µgCulture volume up to 150 ml78711 50 preps
PrepEase Giga Plasmid KitBinding capacity up to 10 mgCulture volume up to 2,000 ml78720 5 preps
Designed to obtain low to high copy plasmid DNA from bacterial cultures.
�� Based on familiar anion-exchange columns �� For routine purification using a well-characterized method �� For use with plasmids from 3 - 10 kb�� Purify from 100 µg up to 10 mg depending on the column size�� Greater than 90% recovery of ultrapure plasmid
PrepEase Plasmid Purification Kits employ a modi-fied alkaline–SDS lysis procedure with patented anion-exchange columns to isolate high purity plasmid DNA from bacterial cell lysates. During the cell lysis step, both chromosomal and plasmid DNA are denatured. Potassium acetate is added to form a neutralized precipitate containing chromo-somal DNA and other cellular components. Plasmid DNA remains in the solution, reverts to its native supercoiled structure, and is then loaded onto an equilibrated PrepEase Column. The plasmid DNA becomes bound to the anion-exchange resin and is then eluted from the column with washing steps. Eluted DNA is precipitated and easily dissolved in TE Buffer or nuclease-free water. The purified plasmids are suitable for use in the most demanding molecu-lar biology applications, including transfection, in vitro transcription, automated or manual sequencing, cloning, hybridization, and PCR.
PrepEase Columns are stable over a wide pH range, from pH 2.5 - 8.5, and can remain in contact with buffers for up to three hours without any change in chromatographic properties. After three hours, nucleic acids will begin to elute at increasingly lower salt concentrations. Normally, the resin remains functional in buffers containing up to 2 M salt, in denaturing agents such as formamide, urea, or in common detergents such as Triton® X-100 and Igepal™ (NP-40 substitute). The protocols are suit-able for purifying most plasmids ranging from 3 to 10 kb. Two protocols are presented, one for purifying high-copy number plasmids (> 20 copies/cell) and the other for purifying low-copy number plasmids (< 20 copies/cell).
Properties of PrepEase ResinPrepEase Resin is a silica-based anion-exchange resin, used for routine separation of different types of nucleic acids. Patented PrepEase Resin forms the basis for the entire line of USB nucleic acid purifica-tion products. The resin consists of hydrophilic, macro-porous silica beads coupled to a methyl-ethylamine functional group. The functional group provides a high overall charge density that permits the negatively charged plasmid DNA to bind with high specificity to the resin.
The beads are uniform in diameter due to a specialized manufacturing process that is rigorously controlled and monitored, and contain particularly large pores. The special properties of PrepEase Resin allow for optimum flow rates through the column and more efficient binding of nucleic acids to the matrix.
Kit components:Each kit includes PrepEase gravity-flow columns and all the necessary reagents for ultrapure plasmid purification.
Includes specialized PrepEase Folded Filters to remove cellular debris from lysates. – Gentle treatment – No shearing of DNA – Fast procedure
Shipping and storage:Shipped ambient. Store at room temperature.
Related products
PrepEase Endotoxin-free Maxi Plasmid Kit78730 10 preps
PrepEase MiniSpin Plasmid Kit78737 250 preps
PrepEase Plasmid 8-well Strip Kit78745 12 strips
PrepEase Plasmid 96-well Plate Kit78751 4 x 96-well plates
PrepEase Quick MiniSpin Plasmid Kit78742 250 preps
PrepEase Plasmid Gravity-Flow Column Kits
141For bulk or alternate pack sizes, email us at [email protected].
78742 250 preps
Very rapid purification
�� Rapid processing of 1 - 3 ml of culture �� For purifying vectors up to 15 kb �� Yields up to 15 μg �� Ideal for direct sequencing �� 50 μl elution volume �� Up to 95% recovery
The PrepEase Quick MiniSpin Plasmid Kit enables rapid, simple purification of plasmid DNA from small volumes of bacterial culture. The kits employs an alkaline lysis procedure and spin columns containing special, patented, anion-exchange membrane filters, which save time by eliminating extra wash and drying steps.
The PrepEase Quick MiniSpin Plasmid Kit protocol is designed for preparation of plasmids ranging up to 15 kb in size, and for high-copy plasmids in particular. Simple modifications enable use of a wide
variety of bacterial strains and growth media, and optimization of elution procedures. For preparation of low-copy plasmids, try the USB PrepEase MiniSpin Plasmid Kit.
Kit components:Includes PrepEase Spin Columns, collection tubes, and all the necessary reagents for rapid plasmid purification.
Shipping and storage:Shipped ambient. Store at room temperature.
PrepEase Quick MiniSpin Plasmid Kit
Purification | Plasmid Purification – Spin Columns
78737 250 preps
High yield purification
�� Process up to 10 ml of culture �� For purifying vectors up to 15 kb �� Yields up to 40 μg �� Ideal for multiple analyses �� 50 μl elution volume �� Up to 95% recovery
The PrepEase MiniSpin Plasmid Kit enables rapid, simple purification of plasmid DNA from small volumes of bacterial culture. The kit employs an al-kaline lysis procedure and patented, anion-exchange membrane filter spin-columns.
The PrepEase MiniSpin Plasmid Kit is designed for preparation of plasmids ranging up to 15 kb in size. Simple modifications of the protocol enable use of a wide variety of bacterial strains and growth media, elimination of problematic nucleases, optimization of wash and elution procedures, and purification of low-copy plasmids. For even faster preparation of high-copy plasmids, try the USB PrepEase Quick MiniSpin Plasmid Kit.
Kit components:Includes PrepEase Spin Columns, collection tubes, and all the necessary reagents for rapid plasmid purification.
Shipping and storage:Shipped ambient. Store at room temperature.
PrepEase MiniSpin Plasmid Kit
PrepEase Standard MiniSpin Column.
PrepEase Quick MiniSpin Column.
Fig. 1. The PrepEase Quick MiniSpin Kit gives high-quality results for auto-mated sequencing. Typical result from plasmid DNA purified using the PrepEase Quick MiniSpin Plasmid Kit, sequenced and analyzed on an ABI PRISM® 3700. Please note the absence of “no calls (N)” far behind the reading length of 700 bp.
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Binding capacity up to 500 µgCulture volume up to 150 ml78730 10 preps
Designed to obtain endotoxin-free plasmids for transfections
�� Based on familiar anion-exchange columns�� Yields plasmid with less than 0.05 EU/µg DNA�� Purify from 500 µg up to 10 mg depending on the column size�� Greater than 90% recovery of ultrapure plasmid
Endotoxins (lipopolysaccharide, LPS) are a major component of gram-negative bacterial cell walls and are extremely potent stimulators of the mammalian immune system. LPS is a common contaminant of plasmid DNA preparations grown in E. coli. The negative charges associated with lipid A and the inner core of LPS cause these molecules to behave like DNA on anion-exchange chromatography resins. Therefore, standard column chromatography procedures typically yield slightly contaminated plasmid DNA.
The PrepEase Endotoxin-Free (EF) Maxi Plasmid Kit utilizes a modified alkaline/SDS lysis procedure to prepare the bacterial cell pellet for plasmid purifica-tion. Both chromosomal and plasmid DNA are dena-tured under alkaline conditions. Potassium acetate is then added to the denatured lysate to form a precipitant of chromosomal DNA and other cellular compounds. The potassium acetate buffer also neu-tralizes the lysate. Plasmid DNA remains in solution and reverts to its native supercoiled structure. The plasmid DNA is bound to an equilibrated PrepEase-EF Column and then eluted after efficient washing of the column. Endotoxins are removed by using a specialized solution, N3-EF Buffer. After precipitation of the eluted DNA, it can easily be dissolved in TE-EF Buffer or EF-Water for further use.
Properties of PrepEase ResinPrepEase-EF Columns contain anion exchange silica resin packed between two inert filter elements. The columns may be used over a wide pH range, from pH 2.5 - 8.5, and can remain in contact with buffers for up to three hours without any change in chromatographic properties. After three hours, nucleic acids will begin to elute at increasingly lower salt concentrations. Normally, the resin remains functional in buffers containing up to 2 M salt. In addition, the resin remains intact in the presence of denaturing agents such as formamide, urea, or common detergents such as Triton X-100 and Igepal (NP-40 substitute).
Kit components:Each kit includes PrepEase Endotoxin-Free Gravity-Flow Columns and all the necessary reagents for ultrapure plasmid purification.
Includes specialized PrepEase Folded Filters/Bottle Top Filters to remove cellular debris from lysates. – Gentle treatment – No shearing of vectors or constructs – Fast procedure
Shipping and storage:Shipped ambient. Store at room temperature.
Related products
Agarose - Separation ≥ 500 bpGenetic Performance Certified™75817 100 gm 250 gm
Agarose - LE 32802 100 gm 250 gm
Ampicillin, Sodium Salt 11259 5 gm 25 gm
Chloramphenicol23660 25 gm
Kanamycin Sulfate 17924 5 gm 25 gm
LB Agar, Ready-Made Powder 75851 250 gm 1 kg
LB Broth, Ready-Made Powder 75852 250 gm 1 kg
Lysozyme 18645 5 gm 10 gm 25 gm
RapidRun™ Agarose Buffer, 20X Solution 77523 1 L 5 L
Streptomycin Sulfate 21865 10 gm 100 gm
TAE Buffer, 10X Solution 75904 1 L 5 L
TBE Buffer, 5X Solution 75891 1 L 5 L
TBE Buffer, 10X Ready-Mixed Powder70454 6 x 200 ml
TE Buffer, 1X Solution75893 10 x 1 ml 100 ml 500 ml
Tetracycline Hydrochloride 22105 25 gm
Water, Nuclease-Free 71786 10 x 1 ml 100 ml
PrepEase Endotoxin-free Maxi Plasmid Kit
Purification | Plasmid Purification – Endotoxin-free Preps
Fig. 1. Typical results with PrepEase plasmid purification products.
143For bulk or alternate pack sizes, email us at [email protected].
Binding capacity up to 20 µgCulture volume up to 5 ml 78745 12 strips
Designed for the isolation of high copy plasmid DNA using 8 well strips
�� Based on familiar silica-membrane technology�� Time-saving purification of multiple samples using 8-well strips�� For use with plasmids up to 15 kb�� Yields up to 20 μg highly pure plasmid DNA from 5 ml of culture: 5 - 15 μg from 1.5 ml of culture�� Process samples by vacuum or robotics
The PrepEase Plasmid 8-well Strip Kit enables rapid purification of plasmid DNA from small volumes of bacterial culture. The kit employs an alkaline lysis procedure, followed by either vacuum filtration, or optionally by robotics, with easy-to-use filter and plasmid binding strips. The plasmid binding strips contain patented anion-exchange membrane filters, allowing highly specific purification of plasmid DNA.
Plasmids ranging in size up to 15 kb may be purified with yields up to 15 μg from 1.5 ml of bacterial culture. The kit is suited for high copy number plasmid purification. Low copy plasmids may be puri-fied by increasing the culture volumes to 5.0 ml. Simple modifications of the protocol enable use of a wide variety of bacterial strains and growth media, elimination of problematic nucleases, and optimization of washes to increase yield.
Use of the PrepEase Plasmid 8-well Strip Kit for the first time requires additional accessories, available separately. For standard vacuum processing, use the PrepEase Vacuum Manifold (PN 78780) and the PrepEase 8-well Strip Starter Kit for Vacuum (PN 78784).
Kit components:Kit includes PrepEase 8-well Binding Strips and all the necessary reagents for high purity plasmid purification.
Includes specialized PrepEase Filter Strips to clear bacterial lysates–replaces time-consuming centrifugation.
Shipping and storage:Shipped ambient. Store at room temperature.
Related products
PrepEase 8-well Strip Starter Kit forVacuumIncludes 2 x 8-well strip holders, dummy strips to close unused rows, and accessories for processing manually or on robotic platform.78784 1 pack
PrepEase Vacuum ManifoldFor use with PrepEase 8-well strips and 96-well plates. Consists of manifold base and lid, spacer set, and a waste tray.78780 1 ea
PrepEase Plasmid 8-well Strip Kit
Purification | Plasmid Purification – Medium Throughput
PrepEase 8-Well Strip Procedure
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Binding capacity up to 20 µgCulture volume up to 5 ml 78751 4 x 96-well plates
Designed for the isolation of high copy plasmid DNA using 96 well plates
�� Based on familiar silica-membrane technology �� Time-saving purification of multiple samples using 96-well plates �� For use with plasmids up to 15 kb �� Yields up to 20 µg highly pure plasmid DNA from 5 ml of culture; 5 - 15 µg from 1.5 ml of culture
The PrepEase Plasmid 96-well Plate Kit enables rapid, simple purification of plasmid DNA from small volumes of bacterial cultures. The kit uses an alkaline lysis procedure, followed by vacuum filtration (and optionally, centrifugation) with filter and plasmid binding plates. The plasmid binding plates contain patented anion-exchange membrane filters, allowing highly specific purification of plasmid DNA. The PrepEase Plasmid 96-well Plate Kit requires a vacuum manifold, such as the PrepEase Vacuum Manifold (PN 78780) or the QiaVac™ 96.
Plasmids ranging up to 15 kb in size may be purified with yields of up to 15 µg from 1.5 ml of bacte-rial culture. The kit is suited for high copy number plasmid purification. Low copy plasmids may be purified by increasing the culture volumes to 5.0 ml. Simple modifications of the protocol enable use of a wide variety of bacterial strains and growth media, elimination of problematic nucleases and optimiza-tion of washes to increase yield.
Kit components:Includes square-well blocks and cover sheets for bacterial cultivation, filter plates, binding plates, wash plates and buffers
Shipping and storage:Shipped ambient. Store at room temperature.
Related products
PrepEase Plasmid 96-well Plate Core KitIncludes filter plates, binding plates, wash plates and buffers. Does not contain square-well blocks for bacterial cultivation.78753 24 x 96-well plates
Related products – Ultrapure reagents
Agarose - Separation ≥ 500 bpGenetic Performance Certified75817 100 gm 250 gm
Agarose - LE 32802 100 gm 250 gm
LB Agar, Ready-Made Powder 75851 250 gm 1 kg
LB Broth, Ready-Made Powder 75852 250 gm 1 kg
Lysozyme 18645 5 gm 10 gm 25 gm
RapidRun Agarose Buffer, 20X Solution 77523 1 L 5 L
TAE Buffer, 10X Solution 75904 1 L 5 L
TBE Buffer, 5X Solution 75891 1 L 5 L
TBE Buffer, 10X Ready-Mixed Powder70454 6 x 200 ml
TE Buffer, 1X Solution75893 10 x 1 ml 100 ml 500 ml
Water, Nuclease-Free 71786 10 x 1 ml 100 ml
Related products – Ultrapure antibiotics
Ampicillin, Sodium Salt 11259 5 gm 25 gm
Chloramphenicol23660 25 gm
Kanamycin Sulfate 17924 5 gm 25 gm
Streptomycin Sulfate 21865 10 gm
Tetracycline Hydrochloride 22105 25 gm
PrepEase Plasmid 96-well Plate Kit
Purification | Plasmid Purification – High Throughput
PrepEase Plasmid Filter Plate
PrepEase Plasmid Binding Plate vacuum
vacuum
highly pure plasmid DNA
screening
restrictiondigests
sequencing
cloning
145For bulk or alternate pack sizes, email us at [email protected].
79220 50 preps
The PrepEase Yeast Plasmid Isolation Kit is designed for the isolation of 2 µ plasmids from either yeast patches on plates or from yeast grown in small liquid culture. The isolation protocol eliminates the need for phenol/chloro form extraction, glass bead disruption and column elution.
The isolated plasmid DNA can be used for PCR amplification, bacterial transformation or for direct sequencing. The ability to sequence directly from the isolated plasmid without the need for bacterial transfor mation or a pre-amplification step and restriction enzyme digestion makes it an efficient method for screening yeast transformants. The kit may also be used to obtain spheroplasts for other analysis.
Direct purificationMinimize the time required to analyze yeast plas-mids, e.g. yeast two-hybrid experiments. The need to shuttle plasmids from yeast to E. coli is eliminated.
Simple protocolAllows the isolation of plasmid from yeast cultures almost as rapidly and readily as from E. coli.
SafeNo organic extraction or mechanical disruption of yeast samples is involved.
Quality resultsYields plasmid DNA suitable for a variety of applica-tions, including direct sequencing, PCR amplifica-tion, and transformation of chemically competent bacterial cells.
Kit components:Enzyme Solution Spheroplast SolutionLysis SolutionPrecipitation Solution
Shipping and storage:Shipped on dry ice. Store at -20°C.
PrepEase Yeast Plasmid Isolation Kit
Binding capacity up to 100 µgCulture volume up to 150 - 500 ml 78722 10 preps
Designed for purification of BACs, PACs, P1s, and other large constructs
�� Based on familiar anion-exchange chromatography�� Purify constructs up to 300 kb�� Yield is typically 10 - 100 µg of BAC vector
The PrepEase BAC Purification Kit employs a modi-fied alkaline–SDS lysis procedure with patented anion-exchange columns to isolate high purity plasmid DNA from bacterial cell lysates. During the cell lysis step, both chromosomal and plasmid DNA are denatured. Potassium acetate is added to form a neutralized precipitate containing chromo-somal DNA and other cellular components. Plasmid DNA remains in the solution, reverts to its native supercoiled structure, and is then loaded onto an equilibrated PrepEase Column. The plasmid DNA becomes bound to the anion-exchange resin and is then eluted from the column with washing steps. Eluted DNA is precipitated and easily dissolved in TE Buffer or nuclease-free water. The purified plasmids are suitable for use in the most demanding molecu-lar biology applications, including transfection, in vitro transcription, automated or manual sequencing, cloning, hybridization, and PCR.
PrepEase Columns contain anion exchange silica resin. The columns are stable over a wide pH range, from pH 2.5 - 8.5, and can remain in contact with buffers for up to three hours without any change in chromatographic properties. After three hours, nucleic acids will begin to elute at increasingly lower salt concentrations. Normally, the resin remains functional in buffers containing up to 2 M salt, in denaturing agents such as formamide, urea, or in common detergents such as Triton X-100 and Igepal (NP-40 substitute).
The BAC purification (low-copy) protocol is suitable for the isolation of low copy plasmids and contains enough buffers and RNase A to perform 10 preps. Typical yields are 10 - 100 µg from 500 ml of fermentation broth depending on copy number and size of constructs.
Kit components:Kit includes PrepEase Gravity-Flow Columns and all the necessary reagents for ultrapure large construct purification.
Specialized PrepEase Folded Filters are used to remove cellular debris from lysates (Fig. 1). – Gentle treatment – No shearing of vectors or constructs – Fast procedure
Shipping and storage:Shipped ambient. Store at room temperature.
PrepEase BAC Purification Kit
Purification | Plasmid Purification – Yeast/BAC
Fig. 2. Typical result using the PrepEase BAC Purification Kit.
Fig. 1. PrepEase Folded Filters are used to remove SDS and cellular debris from bacterial lysates and eliminate a cen-trifugation step.
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Purification | DNA Cleanup/Gel Extraction
Binding capacity up to 15 µg78756 50 preps78757 250 preps
Designed for purifying DNA fragments from agarose gels, and also suited for PCR cleanup
�� Spin columns based on familiar silica-membrane technology�� High recovery rates of up to 90% with low elution volume (15 µl)�� Purify fragments from 50 bp - 10 kb�� Ideal for purifying restriction-digested DNA in gels or cleanup of reaction products
The PrepEase Gel Extraction Kit may be used to purify DNA from agarose gels or to cleanup reaction products. The kit may be used for desalination, re-moval of enzymes, nucleotides, or labeling reagents from sample DNA.
With the PrepEase Cleanup method, DNA binds in the presence of chaotropic salts to the silica mem-brane within the spin column. Contaminants such as salts and soluble macromolecular components are removed with a simple washing step. Pure DNA is eluted under low ionic strength conditions with slightly alkaline buffer.
Typical recovery rates range from 70 - 95% for DNA fragments between 50 - 10,000 bp. The purified DNA may be used directly in applications such as DNA sequencing, PCR, cloning, or probe labeling.
Kit components:Includes PrepEase Spin Columns and all the neces-sary reagents for rapid DNA purification.
Shipping and storage:Shipped ambient. Store at room temperature.
Related products – Ultrapure reagents
Agarose - LE 32802 100 gm 250 gm
Agarose - Low Melt, Separation ≥ 1000 bpGenetic Performance Certified 32830 25 gm 100 gm
Agarose - Low Melt, Separation ≤ 1000 bpGenetic Performance Certified32829 25 gm 100 gm
Ethidium Bromide Drops75816 5 ml
TAE Buffer, 10X Solution 75904 1 L 5 L
Related products – Markers and ladders
DNA Ladder, 1 kb Plus76714 500 µl
DNA Ladder, 100 bp76712 500 µl
PCR Markers, 50 – 2000 bp76710 250 µl
PrepEase Gel Extraction Kits
washing
elution
binding
solubilized gel slice
+ NT Buffer
agarose gel
highly pure DNA fragments
ligation
sequencing
labeling
restrictiondigests
Binding capacity up to 15 µg78758 50 preps78759 250 preps
Designed for the purification of DNA from reactions or for buffer exchange
�� Spin columns based on familiar silica-membrane technology�� High recovery rates up to 90% with low elution volume (15 μl)�� Ideal for cleanup of reaction products�� Binding capacity up to 15 μg�� For DNA fragments 50 – 10,000 bp
The PrepEase DNA Cleanup Kit is designed to purify DNA fragments from enzymatic reactions or for buffer exchange. Thus, this kit can be used to remove
contaminants such as salts, enzymes, nucleotides, labeling reagents, etc. from a DNA sample. This kit can also be used for purification of DNA from Chromatin Immunoprecipitation (ChIP) assays.
The kit provides a simple and convenient way of purifying DNA by selective binding to a silica membrane within the spin column in the presence of a chaotropic salt. The membrane is then washed to remove contaminants such as salts and soluble macromolecules. Pure DNA is then eluted from the membrane with buffer.
Each kit includes PrepEase Cleanup Columns, collection tubes and the necessary reagents for the purification of DNA fragments.
Storage:Ships ambient. Store at room temperature.
PrepEase DNA Cleanup Kits
binding
washing
elution
147For bulk or alternate pack sizes, email us at [email protected].
Purification | Genomic DNA Isolation
78850 50 preps78855 250 preps
Designed for the rapid isolation of genomic DNA
�� Does not use phenol or columns�� Processes large number of samples in a short time�� Optimized for maximal extraction of high molecular weight DNA�� Yields up to 90 μg per prep depending on sample type
The PrepEase Genomic DNA Isolation Kit provides a simple and efficient procedure for the rapid isolation of genomic DNA from various samples, including mouse tails, animal tissues, cultured cells, bacteria, yeast, whole blood, and plant leaves. The procedure does not use phenol or columns and can process a large number of samples in a short time.
This kit has been optimized for maximal extraction of pure high molecular weight genomic DNA. Such genomic DNA is free of protein and ideal for down-stream analyses, including restriction endonuclease digestion, PCR, library construction, Southern blot-ting, cloning, and other applications. The specially formulated Homogenization Buffer not only works well with tissues containing high amounts of nucle-ases, but also with tough fibrous tissues.
Each kit includes all the necessary reagents for the isolation of genomic DNA from a variety of samples. Proteinase K is included for treatment of skin, mouse tails, and other hard tissues.
Storage:Ships ambient. Store individual components as specified.
PrepEase Genomic DNA Isolation Kits
Fig. 1. PCR analysis of transgenic mice. Genomic DNA isolated from mouse tails was analyzed by multiplex PCR to detect three transgenes. PCR products were resolved by electrophoresis in a 1% agarose gel. The unique product for each transgene is shown at the left side of the panel. The mouse genotype is indicated at the bottom of the figure. (Image courtesy of Jianqi Yang, Case Western Reserve University.)
Fig. 3. Genomic DNA isolated from various samples. Genomic DNA was resolved by electrophoresis in a 0.7% agarose gel. Samples are from mouse unless otherwise specified. (Image courtesy of Jianqi Yang, Case Western Reserve University.)
Fig. 2. Southern blotting of C/EBPα alleles in several tissue types. Genomic DNA isolated from various mouse tissues was ana-lyzed using Southern blotting. The unique band for each C/EBPα allele is shown at the left side of the panel; the name of tissue is indicated at the right side. Each lane represents DNA isolated from a single ani-mal. (Image courtesy of Jianqi Yang, Case Western Reserve University.)
148 888-362-2447 | 216-765-5000 | usb.affymetrix.com
78250 20 reactions78200 100 reactions78201 500 reactions78202 2,000 reactions78205 5,000 reactionsCustom formulations by request
�� Conserves PCR samples – 100% recovery of both short and long PCR products �� One tube/one-step PCR cleanup – Add ExoSAP-IT reagent directly to PCR product.�� Eliminates spin columns – Decreases time and expense while increasing yield�� Removes contaminating primers and dNTPs – No interference in downstream applications�� Scalable – Economical for high-throughput purification�� Simple processing – Robotic-friendly; Replaces beads, filtrations, and plates�� Generates less waste than columns
ExoSAP-IT reagent is designed for simple, quick PCR cleanup for downstream applications, such as DNA sequencing or Single Nucleotide Polymor-phism (SNP) analysis. When PCR amplification is complete, any unused dNTPs and primers remain-ing in the PCR product mixture will interfere with these methods. ExoSAP-IT PCR Cleanup removes these contaminants.
ExoSAP-IT reagent is added directly to the PCR product and incubated at 37°C for 15 minutes. After PCR treatment, it is inactivated simply by heating to 80°C for 15 minutes (Fig. 1).
ExoSAP-IT single-step PCR cleanup utilizes two hydrolytic enzymes, Exonuclease I and Shrimp Alkaline Phosphatase (SAP), together in a specially formulated buffer, to remove unwanted dNTPs and primers from PCR products. Exonuclease I removes residual single-stranded primers and any extraneous single-stranded DNA produced in the PCR. SAP re-moves the remaining dNTPs from the PCR mixture.
Rapid PCR product cleanup protocolThe ExoSAP-IT method requires only one pipet-ting step and two incubations. Just add ExoSAP-IT reagent to the PCR product and within 30 minutes sequencing or SNP analysis can be performed.
Simple: single-stepThe method is designed to require a minimum of “hands-on” time. Enzymatic removal of excess primers and unincorporated nucleotides occurs in one easy step by using ExoSAP-IT reagent in a single tube or microtiter well. Only simple pipette transfers are required, therefore, many samples can be pro-cessed at once, either manually or with robotics.
No sample lossUse of ExoSAP-IT reagent eliminates all gel or column purifications, sedimentations, filtrations, beads, and/or magnetic separations(1). There is 100% recovery of both short and long PCR products with ExoSAP-IT reagent.
Achieve high quality data from PCR productsExoSAP-IT reagent may be used as an effective cleanup method prior to fluorescent or radioac-tive DNA sequencing, SNP analysis or any other application requiring a PCR product free of excess nucleotides and primers.
Shipping and storage:Shipped on dry ice. Store at -20°C.
References:1. Dugan, K. A., Lawrence, H. S., Hares, D. R.,
Fisher, C. L. and Budowle B. (2002) J. Forensic Sci 47, 811-818.
2. Hanke, M. and Wink, M. (1994) BioTechniques 17, 858-860.
3. Mu, J., Duan, J., Makova, K., Joy, D., Huynh, C., Branch, O., Li, W. and Su, X. (2002) Nature 418, 323-326.
4. Silva, Jr., W. A., Costa, M. C. R., Valente, V., De Freitas Sousa, J., Pacó-Larson, M. L., Espreafico, E. M., Camargo, S. S., Monteiro, E., De Jesus, A., Holanda, M. A., Zago, M. A., Simpson, A. J. G. and Neto, E. D. (2001) BioTechniques 30, 537-542.
5. Werle, E., Scneider C., Renner, M., Völker, M. and Fiehn, W. (1994) Nucl. Acids Res. 22, 4354-4355.
Related product
HT ExoSAP-IT High-Throughput PCR Product Cleanup78395 480 rctns x 8-tube strip 5,760 rctns x 1 plate (12 x 8-tube strips) 23,040 rctns - 4 plates 1,000 rctns (2 ml) 5,000 rctns (10 ml)
ExoSAP-IT PCR Product Cleanup
Purification | PCR Purification
78761 10 x 96-well plates78762 50 x 96-well plates
Designed for high throughput purification of PCR products using 96-well plates
�� Ultrafiltration technology �� For PCR products ≥150 bp �� Removes contaminating enzymes, salts, primers, and dNTPs �� Process 20 - 300 μl PCR volume per well �� Recover up to 95% depending on length of PCR product in as little as 25 μl
May be performed manually by vacuum, by centrifu-gation, or by using an automated workstation.
Kit components:Includes 96-well plates and buffer for PCR product purification.
Shipping and storage:Shipped ambient. Store at room temperature.
PrepEase PCR Purification 96-Well Plate Kits (Ultrafiltration)
PCR Mixture Post-Amplification
PCR Product
+
Nucleosides
+
Inorganic Phosphate (Pi)
Add ExoSAP-IT®
Excess Primers
Excess dNTPs
37°C, 15 min for treatment80°C, 15 min to inactivate
Fig. 1. Summary of ExoSAP-IT PCR product treatment.
149For bulk or alternate pack sizes, email us at [email protected].
Purification | PCR Purification
78395 480 rctns x 8-tube strip 5,760 rctns x 1 plate (12 x 8-tube strips) 23,040 rctns - 4 plates 1,000 rctns (2 ml) 5,000 rctns (10 ml)
�� One-tube PCR cleanup – Add HT ExoSAP-IT reagent directly to PCR product.�� Exceptional accuracy – Achieve high quality data even with long read lengths (Fig. 1)�� 100% sample recovery – No loss of PCR products regardless of the fragment size (Fig. 2)�� Simple, single-step – Replace multiple steps and wasted time with bead or column use�� High-throughput processing – Even faster time to results with a low viscosity formulation, allowing robotic pipetting�� Removes excess primers and dNTPs – Does not interfere with downstream applications • sequencing • SNP analysis • single base extension • fragment analysis • in vitro transcription�� Scalable – Treat reaction volumes from 5 µl to 5 L�� Convenient packaging – Available in 8-tube strips and 12 x 8-tube strips in a 96-well plate �� Stable at +25°C for 8 hours – Retains full functional activity and at 4°C is stable for one week (Fig. 3)
HT ExoSAP-IT High-Throughput PCR Product Cleanup is an alternative formulation of ExoSAP-IT reagent specifically designed for the unique requirements of high-throughput, automated platform and multi-channel pipettes. HT ExoSAP-IT reagent has both a longer lifetime at higher tempera-tures and a decreased viscosity for robotic pipetting. Like ExoSAP-IT PCR Product Cleanup (PN 78200), HT ExoSAP-IT reagent is a mixture of Exonuclease I and Shrimp Alkaline Phosphatase (SAP) which removes excess primers and dNTPs following a PCR reaction
in a single incubation. HT ExoSAP-IT reagent possesses greater thermal stability and lower viscosity which provides greater ease-of-use in automated 96- and 384-well formats.
HT ExoSAP-IT reagent is added directly to the PCR product and incubated at 37°C for 15 minutes. After PCR treatment, it is inactivated simply by heating to 80°C for 15 minutes. Our high-throughput formula-tion is designed for robotic pipetting and multi-channel pipettes. HT ExoSAP-IT reagent has a lower viscosity than the standard mix and is available in an 8-tube strip format for ease of use.
HT ExoSAP-IT reagent utilizes two hydrolytic enzymes, Exonuclease I and SAP, together in a specially formulated buffer, to remove unwanted dNTPs and primers from PCR products. Exonuclease I removes residual single-stranded primers and any extraneous single-stranded DNA produced in the PCR. SAP removes the remaining dNTPs from the PCR mixture.
Simple: Single-stepHT ExoSAP-IT reagent requires only one pipetting step and two incubations. Just add HT ExoSAP-IT reagent to the PCR product and within 30 minutes sequencing, SNP analysis, single base extension, or fragment analysis can be performed.
Enzymatic removal of excess primers and unincorpo-rated nucleotides occurs in one easy step by using HT ExoSAP-IT reagent in a single-tube, 8-tube strip, or 12 strips in a 96-well low skirted PCR plate. Our new formatting makes this an ideal product for use with robotics.
Exceptional accuracy with HT ExoSAP-IT Achieve high data quality and sequencing accuracy with HT ExoSAP-IT reagent, even with long read lengths. Sequence reads of PCR products treated with HT ExoSAP-IT reagent were on average 50+ bases longer than samples treated with competitor products. Using human genomic DNA, a 1,007 bp fragment was sequenced with no miscalls (Fig. 1). The size limitations associated with alternative PCR cleanup methods are not a factor with HT ExoSAP-IT reagent. Phred 20 values over the entire length of a 970 bp sequence was 822 ± 9 with HT ExoSAP-IT-treated samples and only 776 ± 83 with samples treated with Agencourt® AMPure® XP (see Table 1).
Phred score values of samples processed with HT ExoSAP-IT and AMPure XP BeadsTable 1. HT ExoSAP-IT reagent gives better sequencing results.
HT ExoSAP-IT
AMPure XP beads
Number bases read (Average of 16 samples) 901 850
Total miscalls 0 25Pass rate (%)* 100% 99.7%Phred 20** 822 ± 9 776 ± 83
* Pass rate – Average Phred value greater than 20 bases between 100 bp and 300 bp.
** Phred 20 values over the entire length of sequence (no trim-ming ~970 bp)
No sample lossUse of HT ExoSAP-IT eliminates all gel or column purifications, sedimentations, filtrations, beads, and/or magnetic separations(1). There is 100% recovery of both short and long PCR products with HT ExoSAP-IT High-Throughput PCR Product Cleanup (Fig. 2).
HT ExoSAP-IT-treated PCR product is stableHT ExoSAP-IT reagent is rigorously tested and subject to strict quality control. Fig. 3 demonstrates that no product degradation occurred after storage of HT ExoSAP-IT-treated PCR product for one week at 25°C.
Storage:Shipped on dry ice. Store at -20°C.
References:See references on previous page (PN 78200).
HT ExoSAP-IT High-Throughput PCR Product Cleanup
HT ExoSAP-IT
AMPure XP
3 miscalls
Fig. 1. Sequencing of a 1,007 bp treated PCR product. A 1,007 bp fragment was amplified and treated with HT ExoSAP-IT reagent (above) or Agencourt AMPure XP beads (below) and sequenced. Pherograms revealed no miscalls with HT ExoSAP-IT but three miscalls with Agencourt AMPure XP beads at position 203, 204, and 220.
Fig. 2. Greater recovery of PCR product with HT ExoSAP-IT reagent. Equivalent volumes of PCR product were visualized on an ethid-ium bromide agarose gel and the band volume determined using image analysis. HT-ExoSAP-IT reagent allowed recovery of 100% of the 86 and 103 bp PCR products but AMPure XP beads allowed only 8 and 14% recovery, respectively. Un – Untreated, HT – HT-ExoSAP-IT treated, XP – AMPure XP purified, 100 bp – Affymetrix 100 bp ladder, bands from 100 to 1,000 bp by 100 bp intervals (PN 76712).
Fig. 3. High stability of HT ExoSAP-IT-treated products. A 1,007 bp DNA fragment amplified from genomic DNA with HotStart FideliTaq Master Mix (PN 71156) was treated with HT ExoSAP-IT reagent and stored for seven days at: -20ºC (lane 1), 4ºC (lane 2), or 25ºC (lane 3). Samples were visualized on 1.5% agarose/TAE/ethid-ium bromide gel. Lane MW: DNA Ladder, 1 kb Plus (PN 76714).
1 kb -20°C 4°C 25°C
86 bp 103 bp100 bp Un HT XP Un HT XP
545 bp 1,007 bp100 bp Un HT XP Un HT XP
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Purification | RNA Purification
78878 12 preps
Designed to purify high quality poly(A) RNA from total RNA samples using PrepEase Oligo(dT) Latex Beads.
�� Yields up to 10 µg poly(A) RNA from up to 250 µg total RNA�� Exceptional yield – recover >90% of mRNA from starting total RNA sample�� Fast protocol - process 6 samples in less than 30 minutes�� Excellent purity - purify mRNA from previously isolated total RNA
The PrepEase mRNA MiniSpin Kit produces purified mRNA that is immediately ready for use in demand-ing downstream applications such as qRT-PCR and microarray analysis.
Kit components:Includes PrepEase Oligo(dT) Latex Beads, microfil-ters, and all the necessary reagents for isolation of poly(A) RNA from total RNA samples.
Shipping and storage: Ships ambient. Store PrepEase Oligo(dT) Latex Beads at 4°C. All other kit components may be stored at room temperature.
PrepEase mRNA MiniSpin Kit
Fig. 1. Poly(A) RNA yield vs. total RNA input
Fig. 2. Percent recovery of mRNA (GAPDH) and reduction of ribosomal RNA (rRNA) by PrepEase mRNA MiniSpin Kit from HeLa total RNA. Samples were analyzed by two-step RT-RCR (PN 75780 and 75762).
Binding capacity up to 100 µg78766 50 preps78767 250 preps
Designed for the isolation of total RNA from cells and tissue
�� Spin columns based on familiar silica membrane technology�� Less than 30 minutes hands-on time�� Binding capacity up to 100 µg�� Yields up to 70 µg of high purity DNA-free RNA�� DNase I is included for on-column digestion of contaminating genomic DNA�� Purified RNA may be used directly for RT-PCR, Northerns, microarrays, or cDNA synthesis
Purify total RNA from: Up to 5 x 106 cultured mammalian cells 0.5 - 30 mg mammalian tissue Up to 109 bacterial cells Up to 108 yeast cells Up to 100 µl biological fluids (e.g. serum) RNA cleanup from reaction mixtures
PrepEase RNA Spin Kits are ideal for the isolation of total RNA from mammalian cells, tissues and yeast. The purified RNA has an A260/280 ratio generally ex-ceeding 1.9 and is ready for use in applications such as RT-PCR, primer extension, microarrays, or RNase protection assays. Even biological samples, which are sometimes difficult to process, will yield high quality RNA with the PrepEase RNA Spin Kit. The standard protocol yields up to 70 µg of total RNA per PrepEase RNA Spin Column from up to 5 x 106 cultured cells or 30 mg of tissue.
Kit components:Includes PrepEase Filters to reduce lysate viscosity.
Includes PrepEase RNA Spin Columns, collection tubes, and all the necessary reagents for ultrapure RNA purification.
Shipping and storage:Shipped ambient. Store at room temperature.
PrepEase RNA Spin Kits
cell lysis
homogenization by filtration
binding
DNase I incubationdirectly on thecolumnwashing
elution
Northern blotting primer extensionarray technology
RT-PCR RNase protection
PrepEase RNA and Plant RNA Spin Procedure
151For bulk or alternate pack sizes, email us at [email protected].
Purification | RNA Purification
78871 50 preps
Designed for high quality purification of total RNA and protein from the same sample to allow easy comparison of RNA and protein expression
�� High purity RNA – on column DNase digestion to remove contaminating DNA�� Total protein is prepared in loading buffer, ready for SDS-PAGE analysis�� Straightforward protocol allows efficient workflow
The PrepEase RNA/Protein Spin Kit provides purified total RNA that is immediately ready for sensitive downstream applications such as qRT-PCR and northern blotting. Total protein is eluted in loading buffer for SDS-PAGE analysis.
This kit yields up to 70 μg total RNA and 1.2 mg total protein from a wide array of starting materials, including mammalian tissues and cells, bacteria, yeast, and biological fluids.
Kit components:Includes PrepEase RNA & Protein Spin Columns, filter units, collecting tubes, and all the reagents necessary to isolate both total RNA and protein from the same starting sample.
Shipping and storage: Ships ambient. Store lyophilized rDNase and TCEP at 4°C upon receipt. All other kit components can be stored at room temperature (20-25°C).
PrepEase RNA/Protein Spin Kit
Fig. 1. Procedure overview for the purification of RNA and protein from one sample.
Proteinwash
Proteinprecipitation
RNA washes
RNA elution
rDNase digest
Homogenize sample
Lyse cells
Filter lysate
Bind RNA to column
Preparationof protein
RNA Isolation Protein Isolation
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78776 50 preps78777 250 preps
Designed to remove dye-terminators from sequenc-ing reactions
�� 5 minute protocol�� Based on gel filtration technology�� Ideal for achieving excellent sequencing results�� Spin columns may be stored for long periods
The PrepEase Sequencing Dye Cleanup Kit is designed for easy and rapid separation of excess dye terminators from DNA sequencing reactions. The kit provides spin columns with a specialized gel filtration matrix which binds not only the excess dye terminators, but also impurities such as salts, traces of organic solvents, primers, and free nucleotides. The dye-free sequencing products are eluted from the column and are ready for analysis using standard methods.
Kit components:Includes PrepEase Dye Cleanup Columns and Col-lection Tubes.
Shipping and storage:Shipped ambient. Store at room temperature.
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ExoSAP-IT PCR Product Cleanup78250 20 reactions78200 100 reactions78201 500 reactions78202 2,000 reactions78205 5,000 reactions
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bis-Acrylamide75821 25 gm 50 gm 100 gm 1 kg
RapidGel, 40% Liquid Acrylamide Stock Solution 75848 500 ml
Stop Solution 70724 1200 µl
TBE Buffer, 5X Solution 75891 1 L 5 L
TBE Buffer, 10X Ready-Mixed Powder70454 6 x 200 ml
PrepEase Sequencing Dye Cleanup Kits
Sequencing profile of plasmid DNA after treatment with the PrepEase Sequencing Dye Cleanup Kit.
Purification | Sequencing Dye Cleanup
153For bulk or alternate pack sizes, email us at [email protected].
Purification | Histidine-Tagged Protein Purification Kits
PrepEase Histidine-Tagged Protein Purification Mini Kit - High SpecificityBinding capacity up to 400 µgCulture volume up to 25 ml78791 50 preps
PrepEase Histidine-Tagged Protein Purification Midi Kit - High SpecificityBinding capacity up to 2.5 mgCulture volume up to 150 ml78793 50 preps
PrepEase Histidine-Tagged Protein Purification Maxi Kit - High SpecificityBinding capacity up to 5.0 mgCulture volume up to 300 ml78795 25 preps
Designed for fast, convenient, and high specificity purification of Histidine-tagged proteins
�� High specificity - due to single binding site to Histidine-tagged protein�� Low metal leaching - 5 binding sites to Ni2+
�� High purity > 95%
The PrepEase Protein Purification Kits for High Specificity are designed to purify recombinant polyhistidine-tagged (Histidine-tagged) recombinant proteins expressed in E. coli using immobilized metal ion affinity chromatography (IMAC). The kits may also be used to purify Histidine-tagged proteins from other expression systems such as insect cells, mammalian cells, and yeast. Purification may take place under standard, native conditions, or under denaturing conditions, depending on the solubility of the expressed protein.
PrepEase Protein Purification Kits are based on the use of novel, dry silica-based resin, precharged with Ni2+ ions. The High Specificity Kits utilize TED (tris-carboxymethyl ethylene diamine), a special chelating group, which enables strong and efficient binding of target proteins to the resin. TED is a strong pentadentate metal chelator, which occupies five of the six binding sites in the coordination sphere of the Ni2+ ion. The single remaining binding site can be exchanged with the histidine residues of the recombinant protein (Fig. 1).
Purification methods that utilize other chelators, such as NTA (nitrilotriacetic acid), may lead to metal leaching and/or non-specific binding of untagged proteins to the matrix. NTA chelating groups occupy four of the six binding sites in the coordination sphere of the Ni2+ ion, leaving two sites for protein binding. The additional chelating site of TED mini-mizes metal leaching during purification and reduces non-specific binding of untagged proteins to the ma-trix. This results in high specificity of Histidine-tagged protein binding to the resin, leading to higher purity of the eluted target protein (Fig. 2).
Kit components:Includes PrepEase High Specificity Histidine-tagged Protein Columns and all the necessary reagents for easy protein purification.
Shipping and storage:Shipped ambient. Store at room temperature.
PrepEase Histidine-Tagged Protein Purification Kits - High Specificity
PrepEase Protein Purification Kits�� Novel – Based on dry silica based resin�� Less non-specific binding of contaminating proteins compared to Ni agarose beads�� Easy to elute protein off column�� Utilizes metal (Ni) affinity gravity-flow chromatography�� High stability against chelating and reducing agents�� Room temperature storage
Fig. 1. A: Silica bead bearing Ni2+ bound to TED, pentadentate metal chelator B: One histidine residue of the Histidine-tagged protein binding to the metal ion.
Binding of Histidine-Tagged Protein to PrepEase Ni-TED Resin
KDa
94
67
43
30
20.114.4
M CL F Wash Elution
10 20 50 100 200 250 mM Imidazole
PrepEase Ni-TED
KDa
94
67
43
30
20.114.4
M CL F Wash Elution
10 20 50 100 200 250 mM Imidazole
Ni-NTA AgarosePrepEase Silica-Based Resin Compared to Agarose Beads
Fig. 2. PrepEase High Specificity Ni-TED Resin compared to Ni-NTA Agarose.
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Purification | Histidine-Tagged Protein Purification Kits
PrepEase Histidine-Tagged Protein Purification Mini Kit - High YieldBinding capacity up to 800 μgCulture volume up to 30 ml78801 50 preps
PrepEase Histidine-Tagged Protein Purification Midi Kit - High YieldBinding capacity up to 5 mgCulture volume up to 200 ml78803 50 preps
PrepEase Histidine-Tagged Protein Purification Maxi Kit - High YieldBinding capacity up to 10 mgCulture volume up to 375 ml78805 25 preps
Designed for fast, convenient, and high yield purifi-cation of Histidine-tagged proteins
�� High binding capacity - due to three-binding sites to Histidine-tagged proteins�� 2 times higher binding capacity than PrepEase High Specificity Purification Resin
The PrepEase Protein Purification Kits for High Specificity are designed to purify recombinant polyhistidine-tagged (Histidine-tagged) recombinant proteins expressed in E. coli using immobilized metal ion affinity chromatography (IMAC). The kits may also be used to purify Histidine-tagged proteins from other expression systems such as insect cells, mammalian cells, and yeast. Purification may take place under standard, native conditions, or under denaturing conditions, depending on the solubility of the expressed protein.
PrepEase Protein Purification Kits are based on the use of novel, dry silica-based resin, precharged with Ni2+ ions. Binding of protein is based on the interaction between the polyhistidine tag of the recombinant protein and immobilized Ni2+ ions. The High Yield Kits utilize IDA (iminodiacetic acid), a special chelating group, which enables strong and efficient binding of target proteins to the resin. IDA is a tri-dentate chelator which occupies three of the six binding sites in the coordination sphere of the Ni2+ ion. The remaining three coordination sites can be exchanged with histidine residues of the recombi-nant protein (Fig. 1).
In contrast to traditional IDA matrices, PrepEase Ni-IDA Resin contains an optimized, low density of IDA ligands that are created by a special manufacturing process. The non-saturating surface concentration of IDA eliminates non-specific interactions with contaminating proteins. As a result, PrepEase Ni-IDA Resin allows for higher target protein purity (Fig. 2).
Kit components:Includes PrepEase High Yield Histidine-tagged Protein Columns and all the necessary reagents for easy protein purification
Shipping and storage:Shipped ambient. Store at room temperature.
PrepEase Histidine-Tagged Protein Purification Kits - High Yield
Fig. 1. Structure of PrepEase High Yield Ni-IDA Resin complexed with Ni2+.
PrepEase High Yield Ni-IDA Resin
Fig. 2. PrepEase High Yield Ni-IDA Resin compared to Ni-IDA Agarose.
KDa
9467
43
30
20.1
14.4
M CL Wash Elution
10 20 50 100 250 500 mM Imidazole
PrepEase Ni-IDA Ni-IDA Agarose
KDa9467
43
30
20.1
14.4
M CL Wash Elution
10 20 50 100 250 500 mM Imidazole
PrepEase Silica-Based Resin compared to Agarose Beads
PrepEase Protein Purification Kits�� Novel – Based on dry silica based resin�� Less non-specific binding of contaminating proteins compared to Ni agarose beads�� Easy to elute protein off column�� Utilizes metal (Ni) affinity gravity-flow chromatography�� High stability against chelating and reducing agents�� Room temperature storage
155For bulk or alternate pack sizes, email us at [email protected].
Purification | Histidine-Tagged & GST-Tagged Protein Purification
78796 30 gm
Binding capacity up to 10 mg protein per gm of resin
PrepEase Histidine-tagged High Specificity Purifica-tion Resin is a dry, silica-based resin, precharged with Ni2+ ions. The resin matrix, TED (Tris-carboxy-methyl ethylene diamine), is a pentadentate metal chelator which enables strong and efficient binding of target proteins. Metal leaching and non-specific binding of non-histidine proteins are significantly reduced, resulting in specific, high purity protein.
Shipping and storage:Shipped ambient. Store at room temperature.
PrepEase Histidine-Tagged High Specificity Purification Resin
78806 30 gm 120 gm
Binding capacity up to 20 mg protein per gm of resin
PrepEase Histidine-tagged High Yield Purification Resin is a dry, silica-based resin, precharged with Ni2+ ions. The resin matrix, IDA (iminodiacetic acid), is a tridentate metal chelator which enables strong and efficient binding of target proteins. The PrepEase Ni-IDA Resin has multiple binding sites to Histidine-tagged protein, resulting in higher yields.
Shipping and storage:Shipped ambient. Store at room temperature.
PrepEase Histidine-Tagged High Yield Purification Resin
78820 10 ml 100 ml
Designed for high quality purification of GST-tagged proteins by batch purification, gravity flow, or FPLC™.
�� Yields > 8 mg per 1 ml bed volume�� 60% higher binding capacity compared to Glutathione Sepharose™ 4B�� Flexible – perform batch purification, gravity flow or FPLC�� Easy regeneration – purify more protein over the life of the resin
PrepEase Protein Purification Glutathione Agarose 4B is a medium for single-step purification of glutathione-S-transferase (GST) fusion proteins by affinity chromatography. GST-tagged proteins are purified from cell lysates by their affinity to glutathione which is immobilized on agarose beads. Elution is performed with free glutathione in a non-denaturing, neutral pH buffer. PrepEase Protein Purification Glutathione Agarose 4B is appropriate for purifying GST-tagged proteins by gravity flow chromatography, FPLC, and batch processing.
Kit components:Includes PrepEase Protein Purification Glutathione Agarose 4B for single-step purification of glutathione-S-transferase (GST) fusion proteins by affinity chromatography.
Shipping and storage: Ships ambient. Store at 4°C.
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RNA AnalysismRNA Assay
RNA Transcription
RNA – Reverse Transcription
RNases
RNase Inhibitors
RNA Marker
157For bulk or alternate pack sizes, email us at [email protected].
mRNA AssayPoly(A) Tail-Length Assay Kit 158
RNA TranscriptionE. coli RNA Polymerase Holoenzyme 159Poly(A) Polymerase, Yeast 159T3 RNA Polymerase 160T7 RNA Polymerase 161
RNA - Reverse TranscriptionAMV Reverse Transcriptase 162M-MLV Reverse Transcriptase 162
RNases 163
RNase InhibitorsRNase Inhibitor (Human Placenta) 164RNase Inhibitor (Recombinant) 164
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RNA Analysis Related Products 166
RNA Analysis | Table of Contents
158 888-362-2447 | 216-765-5000 | usb.affymetrix.com
76455 1 kit
Polyadenylation of eukaryotic mRNAs plays a critical role in RNA metabolism including stability, enhanced translation, nuclear export and miRNA mediated gene regulation. The Poly(A) Tail-Length Assay Kit allows you to quickly measure the poly(A) tails of multiple mRNAs from total RNA in only three hours. This kit utilizes a novel G/I extension technique which preserves the 3’ end of RNAs and ensures accurate measurement of poly(A) tail-length. And, all reaction components have been optimized so you get quality data quickly.
�� Measure the poly(A) tail of multiple mRNAs in under 3 hrs using common lab equipment �� Accurate measurement of poly(A) tail length compared to other assays (e.g. northern blotting) �� One kit includes optimized reagents for 5 G/I Tailing, 20 RT and 80 PCR reactions
The Poly(A) Tail-Length Assay Kit uses four key steps to enable poly(A) tail-length determination (Fig. 1).
In Step 1, poly(A) polymerase adds a limited number of guanosine and inosine residues to the 3’-ends of poly(A)-containing RNAs.
In Step 2, the tailed-RNAs are converted to DNA through reverse transcription using the newly added G/I tails as the priming sites.
In Step 3, PCR amplification products are generated using two primer sets. A gene-specific forward and reverse primer set designed upstream of the polyad-enylation site (e.g. the 3’-UTR) is produced as a con-trol for specific detection of the gene-of-interest. The second set of primers uses the gene-specific forward primer and the universal reverse primer provided with the kit to generate a product that includes the poly(A) tails of the gene-of-interest.
Finally, in Step 4, the PCR products are separated on an agarose or polyacrylamide gel (Fig. 2).
Kit components:Kit contains all necessary reagents for 5 G/I Tailing, 20 reverse transcription, and 80 PCR reactions.
Shipping and storage:Shipped on dry ice. Store at -20°C.
References:1. Parker, R. and Song, H. (2004) Nat. Struct. Mol.
Biol. 11 121-127.2. Brodersen, D. E. et al. (2008) Biochim. Biophys.
Acta. 1779, 532-537.
3. Wu, L., Fan J. and Belasco, J. G. (2006) Proc. Natl. Acad. Sci. USA 103, 4034-4039.
4. Izaurralde, E. et al. (2009) RNA 15, 21-32.5. Martin, G. and Keller, W. (1998) RNA 4, 226-230.6. Gauss-Müller, V. et al. (2001) Nucleic Acids Res.
29 E57-7.
Poly(A) Tail-Length Assay Kit
Fig. 2. Comparison of human actin poly(A) tail-length in brain, muscle, liver and HeLa cell. One microgram total RNA and 4 μl diluted RT samples were used in G/I Tailing and PCR reactions, respectively. One half of each PCR reaction (12.5 μl) was analyzed by SYBR Gold-stained 6% non-denaturing polyacrylamide-TBE gel (A), and ethidium bromide-stained 2.5% agarose-TAE gel (B). With RT (+); No RT Negative Control (-); poly(A) tail PCR (A); gene-specific PCR (S); and marker (M).
Fig. 1. Outline of Poly(A) Tail-Length Assay.
RNA Analysis | mRNA Assay
159For bulk or alternate pack sizes, email us at [email protected].
78040Y 100 unitsCustoms by request
Source:E. coli RNA Polymerase Holoenzyme is isolated from the rifampicin-sensitive strain BL21 and is sigma saturated.
Description:E. coli RNA Polymerase is a holoenzyme used for transcription of DNA and the production of labeled RNA for hybridization probes. The polymerase con-sists of 5 subunits designated α, α, β', β and σ. The enzyme is 100% saturated with sigma factor which is required for correct initiation of RNA synthesis on DNA templates containing bacterial and phage promoters.
Application:�� Synthesis of transcripts when it is not possible to transcribe DNA from a vector containing a phage RNA promoter
Properties:Molecular Weight: Approximately 450 kDa. The molecular weight for the subunits are 36.5 (each), 156, 151 and 70 kDa for α, β', β and σ subunits respectively.
Purity:Tested for contaminating non-specific exonucleases, endonucleases and ribonucleases.
Storage buffer:50 mM Tris-HCl (pH 7.5), 250 mM NaCl, 0.1 mM EDTA, 1.0 mM DTT, 50% glycerol.
Assay conditions:The reaction mixture (250 µl) contains 40 mM Tris-HCI (pH 7.5), 150 mM KCI, 10 mM MgCl2, 0.01% Triton X-100, 10 mM DTT, 0.02 µg/µl DNA template, 0.5 mM of each rNTP, and enzyme. Incubation is at 37°C for 10 minutes.
Unit definition:One unit is the amount of enzyme required to cata-lyze the incorporation of 1 nmol of a ribonucleoside triphosphate into RNA in 10 minutes at 37°C.
Concentration:1 unit/µl
Shipping and storage:Shipped on dry ice. Store at -20°C.
References:1. Burgess, R. R. and Travers, A. A. (1970) Fed Proc.
29, 1164-1169.2. Burgess, R. R., Travers, A. A., Dunn, J. J. and Bautz,
E. K. (1969) Nature 221, 43-46.3. Fujiki, H., Palm, P., Zillig, W., Calendar, R. and
Sunshine, M. (1976) Mol. Gen. Genet. 145, 19-22.
E. coli RNA Polymerase Holoenzyme (RNA Nucleotidyl Transferase, E.C.2.7.7.6)
74225Y 10,000 units74225Z 25,000 unitsCustoms by request
Poly(A) Polymerase Reaction Buffer, 5X(Included with each pack size of enzyme)74226 1 ml
Source:E. coli strain containing an overproducing clone of Yeast Poly(A) Polymerase.
Description:Poly(A) Polymerase catalyzes the addition of adeno-sine residues onto the 3'-ends of RNA(1,2). It can be used to add poly(A) tails to RNA in the first step of cloning(3). The reaction requires Mn2+ or Mg2+, ATP as substrate and any RNA containing 3' hydroxyl ter-mini as primer. Longer RNA molecules are somewhat better primers than short oligomers(4). Substitution of cordycepin-5'-triphosphate (3'-dATP) for ATP results in addition of a single 3'-dA residue to the ends of the RNA, a useful technique for labeling RNA at the 3'-end(5)*.
In comparative studies, Yeast Poly(A) Polymerase works more efficiently than E. coli Poly(A) Polym-erase for RNA oligonucleotide-labeling and poly(A) tailing. Shorter incubation times are required for the yeast enzyme and it is found to label both long and short substrates equally well. Poly(A) Polymerase is recommended over T4 RNA Ligase for 3'-end label-ing of long RNA molecules.
*Note: Various modified nucleotides can also be used to label the 3' end of RNA using Yeast Poly(A) Polymerase(5).
Applications:�� Addition of poly(A) tails to RNA�� Labeling the 3' ends of RNA
Purity:Ribonuclease free.
Storage buffer:20 mM Tris-HCl (pH 8.0), 50 mM KCl, 0.5 mM DTT, 50% glycerol.
Assay conditions:25 mM Tris-HCl (pH 7.0), 40 mM KCl, 0.5 mM MnCl2, 0.5 mM EDTA, 0.5 mM DTT, 0.2 mg/ml BSA, 10% glycerol, 3.3 µM radiolabeled ATP, 0.5 mM ATP, 6.5 µg poly(A) (~100 bases), poly(A) polymerase. After incubation at 37°C for 10 minutes, acid insoluble radioactivity is determined.
Unit definition:One unit is the amount of enzyme which incorpo-rates 1 pmol AMP into acid-insoluble material at 37°C in 1 minute.
Concentration:600 units/µl
Functional test:3'-end labeling of a ribonucleotide with cordycepin-5'-triphosphate.
Functionally tested 5X Poly(A) Polymerase Reaction Buffer (1 ml included, PN 74226):100 mM Tris-HCl (pH 7.0), 3.0 mM MnCl2, 0.1 mM EDTA, 1 mM DTT, 500 µg/ml acetylated BSA, 50% glycerol.
Shipping and storage:Shipped on dry ice. Store at -20°C.
References:1. Sippel, A. E. (1973) Eur. J. Biochem. 37, 31-40.2. Edmonds, M. (1982) in The Enzymes, 3rd edition,
ed. P. D. Boyer (Academic Press, New York) 15, 217-244.
3. Gething, M. J., Bye, J., Skehel, J. and Waterfield, M. (1980) Nature 287, 301-306.
4. Sano, H. and Feix, G. (1976) Eur. J. Biochem. 71, 577-583.
5. Martin, G. and Keller, W. (1998) RNA 4, 226-230.
Related products
Adenosine-5'-Triphosphate (ATP), 100 mM Solution77241 25 µmol
Poly(A) Polymerase, Yeast
RNA Analysis | RNA Transcription
160 888-362-2447 | 216-765-5000 | usb.affymetrix.com
Standard concentration, 20 units/µl70051Y 1,000 units70051Z 5,000 unitsHigh concentration, 200 units/µl70204 1,000 units 5,000 units
Source:E. coli strain containing an overproducing clone of T3 RNA Polymerase.
Description:T3 RNA Polymerase has a high specificity for bacteriophage T3 promoter sequences. It is used in the synthesis of large amounts of specific RNA transcripts from vectors containing the T3 promoter. Transcripts may be used as probes in Northern and Southern blots(1), substrates for in vitro translation studies(2), substrates for RNA processing studies(3) and for exon and intron mapping of genomic DNA(1). The enzyme may be used with both radioactively la-beled(4) and hapten-labeled (fluorescein, digoxigenin, biotin, etc.) nucleoside triphosphates(5). T3 RNA Polymerase is also used to generate capped mRNA for expression studies in oocytes and other cells(6).
Applications:�� Production of RNA transcripts�� Production of capped or modified RNA transcripts
Purity:Greater than 90% pure as determined by SDS-PAGE. Tested for contaminating non-specific endonucle-ases, exonucleases and ribonucleases.
Storage buffer:20 mM HPO4 (pH 7.7), 100 mM NaCl, 1 mM EDTA, 10 mM DTT, 1.0% CHAPS, 50% glycerol.
Assay conditions:The reaction mixture contains 1X Transcription Buffer, 0.5 mM each NTP, 1 µCi labeled CTP, 1 µg of T3 promoter containing DNA and T3 RNA Polymerase. Incubation is at 37°C for 15 minutes (50 µl reaction volume).
Unit definition:One unit catalyzes the incorporation of 1 nmol of CMP into polynucleotide fraction bound to a positively charged DE-81 filter paper in 60 minutes at 37°C using a linearized 17 promoter containing 280 nt template.
Concentration:Standard concentration (PN 70051Y/Z): 20 units/µlHigh concentration (PN 70204): 200 units/µl
Functionally tested 10X T7/T3 RNA Polymerase Transcription Buffer:400 mM Tris-HCl (pH 8.0), 60 mM MgCl2, 100 mM DTT, 100 mM NaCl, 20 mM spermidine trihydro-chloride.
Shipping and storage:Shipped on dry ice. Store at -20°C.
References:1. Melton, D. A., Krieg, P. A., Regagliati, M. R.,
Maniatis, T., Zinn, K. and Green, M. R. (1984) Nucl. Acids Res. 12, 7035-7056.
2. Krieg, P. A. and Melton, D. A. (1984) Nucl. Acids Res. 12, 7057-7070.
3. Green, M. R., Maniatis, T. and Melton, D. A. (1983) Cell 32, 681-694.
4. Davanloo, P., Rosenberg, A. H., Dunn, J. J. and Studier, F. W. (1984) Proc. Natl. Acad. Sci. USA 81, 2035-2039.
5. Langer, P. R., Waldrop, A. A. and Ward, D. C. (1981) Proc. Natl. Acad. Sci. USA 78, 6633-6637.
6. Sambrook, J. and Russell, D. W. (2001) Molecular Cloning: A Laboratory Manual, 3rd Edition, Cold Spring Harbor, New York, 9.87-9.88.
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Shrimp Alkaline Phosphatase78390 500 units 1,000 units 5,000 units
T7 RNA PolymeraseStandard Concentration, 20 units/μl70047Y 6,000 unitsHigh Concentration, > 200 units/μl70001Z 30,000 units
Water, Nuclease-Free71786 10 x 1 ml 100 ml 500 ml 1 L 5 L
Related products – Nucleotides
Adenosine-5'-Triphosphate (ATP), 100 mM Solution77241 25 µmol
Guanosine-5'-Triphosphate (GTP), 100 mM Solution77243 25 µmol
NTPs, Set of Four, (ATP, CTP, GTP, UTP), 100 mM Solutions, 4 x 25 µmol77245 4 x 25 µmol
T3 RNA Polymerase (RNA Nucleotidyl Transferase, E.C.2.7.7.6)
RNA Analysis | RNA Transcription
161For bulk or alternate pack sizes, email us at [email protected].
Standard concentration, 20 units/µl70047Y 6,000 unitsHigh concentration, >200 units/µl70001Z 30,000 unitsCustoms by request
Source:E. coli strain containing an overproducing clone of T7 RNA Polymerase(1).
Description:T7 RNA Polymerase is a single-subunit enzyme produced by bacteriophage T7(2). It is highly specific for T7 promoter and terminator sequences(2,3). It has been widely used for the rapid synthesis in vitro of specific RNAs. These transcripts can be used directly as substrates for studies of RNA structure or me-tabolism. If the transcripts are suitably labeled, they can also be used as sensitive hybridization probes. T7 RNA Polymerase, along with chain-terminating nucleoside triphosphates(4), have also been used for the direct sequencing of DNA. T7 RNA Polymerase is also used to generate capped mRNA for expression studies in oocytes and other cells(5).
Applications:�� Production of RNA transcripts for hybridization probes�� Production of large amounts of discrete sized RNA�� Production of capped or modified RNA transcripts
Properties:Molecular Weight: 98.8 kDaOptimum pH: 7.7 - 8.3Optimum Temperature: 37°CRequirement for Divalent Cation: Mg2+ (optimal
concentration is 20 mM)Michaelis Constants(2): 40 µM ATP, 160 µM GTP,
60 µM UTP, 80 µM CTP
Purity:Greater than 99% pure as determined by SDS-PAGE. Tested for contaminating nonspecific endonucleases, exonucleases and ribonucleases.
Storage buffer:20 mM HPO4, pH 7.7, 100 mM NaCl, 1 mM EDTA, 10 mM DTT, 1% CHAPS and 50% glycerol.
Assay conditions:The reaction mixture contains 1X Transcription Buffer, 0.5 mM each NTP, 1 µCi labeled CTP, 1 µg of T7 promoter containing DNA and T7 RNA Polymerase. Incubation is at 37°C for 15 minutes (50 µl reaction volume).
Unit definition:One unit catalyzes the incorporation of 1 nmol of CMP into polynucleotide fraction bound to a positively charged DE-81 filter paper in 60 minutes at 37°C using a linearized T7 promoter containing 280 nt template.
Concentration:Standard concentration (PN 70047Y): 20 units/µlHigh concentration (PN 70001Z): >200 units/µl
Functional test:Transcription from plasmid containing T7 promoter; >10 µg of RNA can be produced from 1 µg of supercoiled template.
Functionally tested 10X T7/T3 RNA Polymerase Transcription Buffer:400 mM Tris-HCl, pH 8.0, 60 mM MgCl2, 100 mM DTT, 100 mM NaCl, 20 mM spermidine trihydro-chloride.
Shipping and storage:Shipped on dry ice. Store at -20°C.
References:1. Tabor, S. and Richardson, C. C. (1985) Proc. Natl.
Acad. Sci. USA 82, 1074-1078.2. Chamberlin, M. and Ring J. (1973) J. Biol. Chem.
248, 2235-2244, 2245-2250.3. Chamberlin, M. and Ryan, T. (1982) in The
Enzymes, 3rd edition, ed. P. D. Boyer (Academic Press, New York.) 15, 87-108.
4. Axelrod, V. D. and Kramer, F. R. (1985) Biochemistry 24, 5716-5723.
5. Sambrook, J. and Russell, D. W. (2001) Molecular Cloning: A Laboratory Manual, 3rd Edition, Cold Spring Harbor, New York, 9.87-9.88.
6. Ausubel, F. M., Brent, R., Kingston, R. E., Moore, D. D., Seidman, J. G., Smith, J. A. and Struhl, K. (1998) Current Protocols in Molecular Biology (John Wiley and Sons, Inc.).
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RT-PCR Master Mix (2X)78370 100 reactions
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TAE Buffer, 10X Solution 75904 1 L 5 L
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T7 RNA Polymerase (RNA Nucleotidyl Transferase, E.C.2.7.7.6)
RNA Analysis | RNA Transcription
162 888-362-2447 | 216-765-5000 | usb.affymetrix.com
70041Y 200 units70041Z 1,000 unitsCustoms by request
Source:Purified virion of Avian myeloblastosis virus.
Description:AMV Reverse Transcriptase catalyzes the polymeriza-tion of DNA using template DNA, RNA or RNA:DNA hybrids(1). It consists of two polypeptide chains, one of which contains a 5'→3' polymerase activity and the other an RNase H activity. Reverse transcriptase requires Mg2+ or Mn2+ and is used for first strand synthesis of complementary DNA (cDNA) from mRNA template(2). Under proper conditions, high yields of full-length cDNA can be obtained with AMV Reverse Transcriptase(3). AMV Reverse Transcriptase is also useful in the amplification of RNA(4).
Applications:�� Synthesis of cDNA for PCR, cloning and hybridization probes�� Filling-in and labeling the 3' termini of DNA with 5' protruding ends�� RNA sequencing�� Amplification of RNA(4)
�� Primer extension assays
Properties:Molecular Weight: 157 kDaInactivation: 75°C for 10 minutes or by adding 2 µl
of 0.5 M EDTA for a 50 µl reaction.
Purity:Tested for contaminating endonucleases, exonucle-ases and ribonucleases.
Storage buffer:200 mM potassium phosphate (pH 7.2), 2 mM DTT, 0.2% Triton X-100, 50% glycerol.
Assay conditions:The reaction mixture (100 µl) contains 50 mM Tris-HCI (pH 8.3), 6 mM MgCl2, 40 mM KCI, 0.5 mM radiolabeled TTP, 0.4 mM poly(rA)n:oligo(dT)50. Incubation is at 37°C for 10 minutes.
Unit definition:One unit is the amount of enzyme required to incor-porate 1 nmol of radiolabeled nucleotide into acid insoluble product in 10 minutes at 37°C.
Concentration:15 units/µl
Functional test:RT-PCR amplification on generation of 0.5 kb and 1.5 kb RT-PCR products.
Functionally tested 5X AMV RT Reaction Buffer (1 ml included):250 mM Tris-HCl (pH 8.3), 40 mM MgCl2, 250 mM NaCl, 5 mM DTT
Shipping and storage:Shipped on dry ice. Store at -20°C. Avoid freeze-thaw cycles, which result in loss of enzyme activity.
References:1. Kacian, D. L. (1977) in Methods in Virology, eds.
K. Maramorosch and H. Koprowski 6, 143-184.2. Goodman, H. M. and MacDonald, R. J. (1979)
Methods Enzymol. 68, 75-90.3. Berger, S. L., Wallace, D. M., Puskas, R. S. and
Eschenfeldt, W. H. (1983) Biochemistry 22, 2365-2372.
4. PCR Protocols: A Guide to Methods and Applications (1990) eds. M. A. Innis, D. H. Gelfand, and John J. Sni, (Academic Press, New York), 23.
AMV Reverse Transcriptase
78306 25,000 units 100,000 unitsCustoms by request
M-MLV Reaction Buffer, 5X(Included with each pack size of enzyme)71505 1 ml
Source:E. coli strain containing an overproducing clone of M-MLV Reverse Transcriptase.
Description:M-MLV Reverse Transcriptase catalyzes the polymerization of DNA using template DNA, RNA or RNA:DNA hybrids(1). Full-length copies of large mRNAs, >10 kb, may be synthesized. M-MLV Reverse Transcriptase has a much lower RNase H activity than AMV Reverse Transcriptase resulting in high yields of full length cDNA(2). This makes M-MLV Reverse Transcriptase very useful in cDNA synthesis and RT-PCR.
For added convenience and optimal RT-PCR, try the USB RT-PCR Kits (PN 78350 and 78355) and RT-PCR Master Mixes (PN 71185 and 78370).
Applications:�� RT-PCR�� Synthesis of first strand cDNA for PCR, cloning and hybridization probes�� Filling-in and labeling the 3' termini of DNA with 5' protruding ends�� Amplification of RNA�� Primer extension assays
Properties:Molecular Weight: 71 kDa (monomeric).Inhibitors: Polyamines, phosphate, pyrophosphates
and lithium chloride(3,4).Inactivation: 75°C for 10 minutes or by adding 2 µl
of 0.5 M EDTA for a 50 µl reaction.
Purity:Greater than 90% pure as determined by SDS-PAGE. Tested for contaminating endonucleases, exonucle-ases and ribonucleases.
Storage buffer:20 mM Tris-HCl (pH 7.5), 0.1 M NaCl, 0.1 mM EDTA, 1 mM DTT, 0.01% Igepal CA-630, 50% glycerol.
Assay conditions:The reaction mixture (25 µl) contains 50 mM Tris-HCI (pH 8.3), 40 mM KCl, 6 mM MgCl2, 1 mM DTT, 0.4 mM poly(rA)-oligo(dT)12 - 18, 0.5 mM radiolabeled TTP and 0.1 - 0.5 units enzyme. Incubation is at 37°C for 10 minutes.
Unit definition:One unit is that amount of enzyme required to incorporate 1 nmol of deoxynucleotide into DE-81 filter-binding material in 10 minutes at 37°C using poly (rA)-oligo(dT)12-18 as template-primer.
Concentration:200 units/µl
Functional test:RT-PCR amplification on generation of 0.5 kb and 1.5 kb RT-PCR products.
Functionally tested 5X M-MLV Reaction Buffer (1 ml included, PN 71505):250 mM Tris-HCl (pH 8.3), 375 mM KCl, 15 mM MgCl2, 50 mM DTT.
Shipping and storage:Shipped on dry ice. Store at -20°C.
References:1. Roth, M. J., Tanese, N. and Goff, S. P. (1985)
J. Biol. Chem. 260, 9326-9335.2. Sambrook, J. and Russell, D. W. (2001) Molecular
Cloning: A Laboratory Manual, 3rd Edition, Cold Spring Harbor, New York, 8.48.
3. Gerard, G. F. and D’Alessio, J. M. (1993) Methods In Molecular Biology 16, Humana Press, NJ, 73-93.
4. Sambrook, J. and Russell, D. W. (2001) Molecular Cloning: A Laboratory Manual, 3rd Edition, Cold Spring Harbor, New York, A8.16.
M-MLV Reverse Transcriptase (E.C.2.7.7.49)
RNA Analysis | RNA—Reverse Transcription
163For bulk or alternate pack sizes, email us at [email protected].
Prod. No. Product Description Size
27-0330-02 RNase I 1 gm See page 79 RNase I catalyzes the hydrolysis of 3'→5' phosphodiester bonds of RNA with the formation of oligoribonucleotides terminating in pyrimidine 2'→3' cyclic phosphate.
27-0323-01 RNase I ‘A’, Powder 100 mg See page 79 RNase I ‘A’ specifically cleaves single-stranded RNA at 3' phosphate linkages of pyrimidine residues leaving pyrimidine 3' phosphates and oligonucleotides with terminal pyrimidine 3' phosphates.
78020Y RNase A, Lyophilized 100 mg See page 79 RNase A specifically cleaves single-stranded RNA at 3' phosphate linkages of pyrimidine residues leaving pyrimidine 3' phosphates and oligonucleotides with terminal pyrimidine 3' phosphates.
70194Y/Z RNase A, Molecular Biology Grade Solution 1 mg See page 80 RNase A specifically cleaves single-stranded RNA at 3' 5 mg phosphate linkages of pyrimidine residues leaving pyrimidine 3' phosphates and oligonucleotides with terminal pyrimidine 3' phosphates.
70054Y/Z RNase H 250 units See page 80 E. coli RNase H is an endoribonuclease that degrades the 1,000 units RNA portion of DNA:RNA hybrids.
RNases
RNA Analysis | RNases
Customized solutions
�� Our experts in custom and OEM reagents will work with you at your site and from our headquarters�� Our over 3,000 catalog products can be tailored to meet your specific needs�� Custom fill sizes, concentrations, and formulations�� Volume fills from microliter to liters�� Weight fills of milligrams to kilograms�� Customer-specific quality control assays�� Liquid conversion to powder in pre-weighed packaging
Most of our products can be tailored to meet your specifications. With our high standards, ISO certification, and scalable purchasing, we are well positioned to offer you a wide range of customized quality products.
To learn more, visit usb.affymetrix.com/bulk.
164 888-362-2447 | 216-765-5000 | usb.affymetrix.com
71570 5,000 unitsCustoms by request
Source:Human placenta
Description:Human Placental RNase Inhibitor (HPRI) binds and inhibits a broad range of eukaryotic RNases, includ-ing RNase A, RNase B, RNase C and human placen-tal RNase. HPRI is purified from human placenta by affinity chromatography on an immobilized RNase A column. It forms a 1:1 complex with RNase A and inhibits RNase activity non-competitively (Ki = 3 x 1010 M). It can be added directly to reaction mixtures containing RNA. HPRI differs from other competitive inhibitors in that it can be easily removed from reac-tion mixtures by phenol extraction.
RNase Inhibitor is not effective against RNase 1, RNase T1, S1 Nuclease, RNase H or RNase from Aspergillus. To maintain activity, HPRI requires a minimum of 1 mM DTT and is active over a broad pH range (pH 5.5-9.0).
Applications:�� cDNA synthesis(2)
�� In vitro translation(3)
�� In vitro transcription(4)
�� Polysome isolation�� RT-PCR�� Isolation of mammalian cell samples that contain a mRNA-protein complex(3)
�� Separation and identification of specific ribonuclease activities(5)
Properties:Activators: Requires DTT for activityInhibitors: RNases will recover activity after
inactivation at temperatures greater than 55°C or in 7 M urea.
Purity:Tested for contaminating non-specific endonu-cleases, exonucleases, ribonucleases and latent ribonucleases.
Storage buffer:20 mM HEPES-KOH (pH 7.6), 50 mM KCl, 8 mM DTT, 50% glycerol.
Unit definition:One unit inhibits the activity of 5 ng of RNase A by 50%(1). This inhibitor activity is determined by its ability to inhibit the hydrolysis of cyclic 2' 3'-CMP by RNase A.
Concentration:≥ 20 units/µl
Shipping and storage:Shipped on dry ice. Store at -20°C. Avoid repeated freeze/thaw cycles.
References:1. Blackburn, P. (1979) J. Biol. Chem. 254, 12484-
12487.2. de Martynoff, G., Pays, E. and Vassart, G. (1980)
Biochem. Biophys. Res. Commun. 93, 645-653.3. Scheele, G. and Blackburn, P. (1979) Proc. Natl.
Acad. Sci. USA 76, 1898-1902.4. Nielson, D. A. and Shapiro, D. J. (1986) Nucl. Acids
Res. 14, 5936.5. Eichler, D. C., Tatar, T. F. and Lasater, L. S. (1981)
Biochem. Biophys. Res. Commun. 101, 396-403.
RNase Inhibitor (Human Placenta)
71571 5,000 unitsCustoms by request
Source:E. coli strain containing an overproducing clone of human placenta ribonuclease inhibitor.
Description:Recombinant Human Placental RNase Inhibitor (rHPRI) binds and inhibits a broad range of eukary-otic RNases, including RNase A, RNase B, RNase C and human placental RNase. HPRI forms a 1:1 complex with RNase A and inhibits RNase activity non-competitively (Ki = 3 x 1010 M). It can be added directly to reaction mixtures containing RNA. HPRI differs from other competitive inhibitors in that it can be easily removed from reaction mixtures by phenol extraction.
RNase Inhibitor is not effective against RNase 1, RNase T1, S1 Nuclease, RNase H or RNase from Aspergillus. To maintain activity, HPRI requires a minimum of 1 mM DTT and is active over a broad pH range (pH 5.5-9.0).
Applications:�� cDNA synthesis(2)
�� In vitro translation(3)
�� In vitro transcription(4)
�� Polysome isolation�� RT-PCR�� Isolation of mammalian cell samples that contain a mRNA-protein complex(3)
�� Separation and identification of specific ribonuclease activities(5)
Properties:Molecular Weight: 50 kDaIsoelectric Point: 4.7Activators: DTTInhibitors: RNases will recover activity after
inactivation at temperatures greater than 55°C or in 7 M urea.
Purity:Greater than 90% pure as determined by SDS-PAGE. Tested for contaminating non-specific endonucle-ases, exonucleases and ribonucleases.
Storage buffer:20 mM HEPES-KOH (pH 7.6), 50 mM KCl, 8 mM DTT, 50% glycerol.
Unit definition:One unit inhibits the activity of 5 ng of RNase A by 50%(1).
Concentration:40 units/µl
Shipping and storage:Shipped on dry ice. Store at -20°C. Avoid repeated freeze/thaw cycles.
References:1. Blackburn, P. (1979) J. Biol. Chem. 254, 12484-
12487.2. de Martynoff, G., Pays, E. and Vassart, G. (1980)
Biochem. Biophys. Res. Commun. 93, 645-653.3. Scheele, G. and Blackburn, P. (1979) Proc. Natl.
Acad. Sci. USA 76, 1898-1902.4. Nielson, D. A. and Shapiro, D. J. (1986) Nucl. Acids
Res. 14, 5936.5. Eichler, D. C., Tatar, T. F. and Lasater, L. S. (1981)
Biochem. Biophys. Res. Commun. 101, 396-403.
RNase Inhibitor (Recombinant)
RNA Analysis | RNase Inhibitors
165For bulk or alternate pack sizes, email us at [email protected].
76410 100 µl
Description:The Low Molecular Weight Marker is a set of 14 bands between 10 - 100 nucleotides, designed for size determination of single-stranded oligonucleotide fragments.
The marker is suitable for use in either polyacryla-mide or agarose gel electrophoresis. The recom-mended concentration for agarose is 2.5% and for polyacrylamide is 12 - 15%. This marker should be diluted with TE Buffer and the supplied Gel Loading Buffer to the desired loading concentration.
The Low Molecular Weight Marker can be used to generate a 5'-end radiolabeled marker for size comparison of small, single-stranded oligonucleotide fragments such as miRNA, siRNA, tRNA, snRNA, snoRNA, or ssDNA. The Low Molecular Weight Marker is also useful when the molecule of interest is less than 100 nucleotides in length. This marker is not designed for precise quantification of nucleic acid mass and should only be used as a size marker.
2X Formamide Loading Buffer and 6X Glycerol Load-ing Buffer are included for convenience to prepare both the marker and samples for gel electrophoresis.
Fragment sizes: 100 nt 40 nt 90 nt 35 nt 80 nt 30 nt 70 nt 25 nt 60 nt 20 nt 50 nt 15 nt 45 nt 10 nt
Functional test: Meets or exceeds criteria for 5'-end labeling with isotope and polyacrylamide gel electrophoresis.
Components:Low Molecular Weight Marker, 100 µl (50 ng/µl each
fragment)2X Formamide Loading Buffer6X Glycerol Loading Buffer
2X Formamide Loading Buffer (1 ml included): Consists of bromophenol blue, xylene cyanol FF, EDTA and formamide.
6X Glycerol Loading Buffer (1 ml included): Consists of orange G, xylene cyanol FF and glycerol.
Shipping and storage:Shipped on ice. Store at 4°C or at -20°C for long term storage.
To use:1. Mix 2 µl (100 ng each fragment) of Marker with
2 µl of 2X Formamide Loading Buffer, heat at 95°C for 3 minutes, quick cool on ice, and load the entire volume per lane on polyacrylamide gel.
2. Mix 5 µl (250 ng each fragment) of Marker with 1 µl of 6X Glycerol Loading Buffer and load the entire volume per lane on agarose gel.
3. Visualize with dyes for detecting single-stranded oligonucleotide fragments.
4. For 5'-end labeling reaction, use 1 µl (50 ng each fragment) of Marker with OptiKinase and [γ-32P]-ATP or [γ-33P]-ATP.
Related products – miRNA Detection
OptiKinase78334X 500 units78334Y 1,000 units78334Z 2,500 units
Related products – UItrapure electrophoresis reagents
Ammonium Persulfate 76322 100 gm
Glycerol Tolerant Gel Buffer, 20X Solution75827 1 L
RapidGel, 40% Liquid Acrylamide Stock Solution75848 500 ml
TAE Buffer, 10X Solution 75904 1 L 5 L
TBE Buffer, 5X Solution75891 1 L 5 L
TE Buffer, 1X Solution75893 10 x 1 ml 100 ml 500 ml
TEMED76320 100 ml 100 gm 500 gm
Low Molecular Weight Marker, 10 – 100 nt
Fig. 1. Low Molecular Weight Marker on 12-15% UREA-polyacrylamide gel. A. 50 ng of SYBR® Gold stained Marker. B. 0.5 ng of 5' end labeled Marker with [γ-33P]-ATP and OptiKinaseTM. C. 0.05 ng of 5' end labeled Marker with [γ-32P]-ATP and OptiKinase.
100908070
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A B Cnt nt nt
RNA Analysis | RNA Marker
166 888-362-2447 | 216-765-5000 | usb.affymetrix.com
Prod. No. Product Description Size
76410 Low Molecular Weight Marker, 10-100 nt 100 µl
RNA Marker
Prod. No. Product Description [conc] Size
77241 ATP 100 mM 25 μmol (250 μl) Adenosine-5'-Triphosphate Ultrapure
77243 GTP 100 mM 25 μmol (250 μl) Guanosine-5'-Triphosphate Ultrapure
77245 NTPs, Set of Four (ATP, CTP, GTP, UTP) 100 mM 4 x 25 μmol (250 μl) Nucleoside Triphosphates 4 NTPs per pack (pk) Ultrapure 1 pk
Related Products
Oligo p(dT)
Prod. No. Product Description Size
19817 Oligo p(dT)12-18 Sodium Salt 5 units 25 units 100 units
RNA Electrophoresis
Prod. No. Product Description Size
76322 Ammonium Persulfate (APS) 10 gm 100 gm 1 kg
75827 Glycerol Tolerant Gel (GTG) Buffer, 20X Solution 1 L
75848 RapidGel, 40% Liquid Acrylamide Stock Solution 500 ml
75891 TBE Buffer, 5X Solution 1 L 5 L
76320 TEMED 100 ml 100 gm 500 gm
75826 Urea 1 kg 5 kg
Ribonucleotides (NTPs), Sodium Salt Solutions
RNA Analysis | Related Products
167For bulk or alternate pack sizes, email us at [email protected].
Related Products continued…
RNA Ligation
Prod. No. Product Description Size
21245 T4 RNA Ligase 600 units
RNA Purification
Prod. No. Product Description Size
78766 PrepEase RNA Spin Kit 50 preps
78878 PrepEase mRNA MiniSpin Kit 12 preps
RNAse-Free Buffers & Solutions
Prod. No. Product Description Size
75901 Ammonium Acetate (NH4OAc), 5 M Solution 100 ml 500 ml
15694 EDTA, 0.5 M Solution 100 ml 500 ml
78641 Magnesium Chloride (MgCl2), 1 M Solution 10 x 1 ml 100 ml
75896 Potassium Chloride (KCl), 2 M Solution 100 ml
75888 Sodium Chloride (NaCl), 5 M Solution 100 ml 500 ml 1 L
22637 Tris/HCl, 1 M Solution, pH 7.0 100 ml 500 ml 1 L
22638 Tris/HCl, 1 M Solution, pH 8.0 100 ml 500 ml 1 L
71786 Water, Nuclease-Free 10 x 1 ml 100 ml 500 ml 1 L 5 L
70783 Water, RNase-Free, DEPC Treated 25 ml 10 x 1 ml 100 ml 500 ml 1 L
RNA Analysis | Related Products
DNA SequencingNGS Library Prep
Thermo Sequenase™ Kits
Sequenase Enzyme & Kits
Sequencing Primers
Sequencing Tips
169For bulk or alternate pack sizes, email us at [email protected].
DNA Sequencing | Table of Contents
Next Generation Sequencing (NGS) Library PrepPrep2Seq™ DNA Library Prep Kit for
Illumina platform 170Prep2Seq Multiplex Oligo Adapters for
Illumina platform 171
Thermo Sequenase KitsThermo Sequenase Cycle Sequencing Kit 172Thermo Sequenase Dye Primer Manual Cycle
Sequencing Kit 173
Sequenase Enzyme & KitsSequenase Version 2.0 DNA Polymerase 174Sequenase Version 2.0 DNA Sequencing Kit 175Sequenase PCR Product Sequencing Kit 176Sequenase Quick Denature Plasmid DNA
Sequencing Kit 177
Sequencing PrimersPrimers for DNA Sequencing 178
Sequencing TipsDetermining Primer Concentration 178DNA Sequencing Gel Formulations 178Use of Terminal Deoxynucleotidyl Transferase
(TdT) to Resolve Bands Across Four Lanes (BAFLs) 179
Relief from Compression Artifacts in DNA Sequencing Using Formamide Gels 180
Prevent Weak Sequencing Band Intensities by Using Pyrophosphatase 181
170 888-362-2447 | 216-765-5000 | usb.affymetrix.com
79900 10 reactions 20 reactions
Generate high quality next generation sequencing (NGS) libraries suitable for sequencing on Illumina platforms in less time with Prep2Seq DNA Library Prep Kit.
DNA library prep is critical to the precision and accuracy of NGS. The Prep2Seq DNA Library Prep Kit comes with all of the reagents necessary for end repair, A-tailing, and ligation, and can be completed in less than two hours.
Our kit is fast, efficient, and optimized. It efficiently converts 3’ and 5’ overhangs into 3’ A-overhang ends for ligation to T-tailed DNA adapters and reduces the potential for bias with our superior A-tailing and ligation steps. We offer significant improvement in the quality of sequence-ready libraries by eliminating chimeras and reducing side reaction fragments.
The Prep2Seq DNA Library Prep Kit combines end re-pair and A-tailing steps, decreasing the total number of steps to achieve quality NGS library preparation. Only one cleanup step is required to attain peak performance on the Illumina sequencing platform and only 100 ng of input is needed.
Full kit offering�� Single kit that includes all reagents for sample prep in a slimmed down protocol.�� 24 bar code adapter sequences compatible with Illumina technology are sold separately:
Prep2SeqMultiplexOligoAdaptersforIllumina - Index Adapter Set I–adapter sequences 1-12 - Index Adapter Set II–adapter sequences 13-27
Time savings with a more efficient method for NGS sample prep�� Improved A-tailing reaction with proprietary enzymes, cloning know-how, and optimized buffers.�� By combining the end repair and A-tailing steps, only one cleanup step is needed compared to the two or three steps in most other kits (saving up to two and a half hours in the lab).
The Prep2Seq DNA Library Prep Kit offers the highest adapter ligation efficiency available. By combining end repair and A-tailing, the Prep2Seq DNALibrary Prep Kit is able to reduce total NGS library prep time to 110 minutes and limit bead cleanup to a single step.
Rigorous comparison studies to competitor kits show that the Prep2Seq DNA Library Prep Kit generates impressive sequencing data.
Prep2Seq™ DNA Library Prep Kit for Illumina platform
DNA Sequencing | Next Generation Sequencing Library Prep
Illumina TruSeq™ Nano
140 min
NEB Next® Ultra
65 min
USB Prep2Seq™
40 minHands-on Lab Time
Walk-Away Time
USB - 110 total min
NEB - 155 total min
Illumina - 210 total minIllumina TruSeq Nano
70 min
NEB Next Ultra
90 min
USB Prep2Seq
70 min
Time Comparison
from sheared DNA to size selection
Table 1. Validated performance of the Prep2Seq DNA Library Prep Kit compared to competition.
USB® Prep2Seq™ Illumina® TruSeq™ NEBNext® Ultra™
Total Reads 170,507,429 171,139,925 176,073,965
Mapped reads to a single locus 142,223,440 143,567,898 147,688,157
% Mapped reads to a single locus 83.41% 83.89% 83.88%
Mapped reads to multiple loci 12,270,818 11,794,099 12,266,301
% Mapped reads to multiple loci 7.20% 6.89% 6.97%
Unmapped reads 16,013,171 15,777,928 16,119,507
% Unmapped reads 9.39% 9.22% 9.15%
The HiSeq® 2500 raw reads were aligned to the human genome hg19 release using Bowtie, allowing 2 mismatches for the entire read length. Mapped reads aligning to a single locus, mapped reads aligning to multiple loci, as well as unmapped reads from the 3 libraries are summarized in Table 1.
Table 2. GC content present in mapped and unmapped reads for the 3 library prep methods.
USB® Prep2Seq™ Illumina® TruSeq™ NEBNext® Ultra™
% GC: Mapped reads to a single locus 42.27% 40.01% 41.10%
% GC: Mapped reads to multiple loci 43.53% 41.65% 42.55%
% GC: Unmapped reads 40.90% 38.79% 40.64%
In order to determine if the end-repair and adaptor ligation may have contributed to some library bias we compared the overall GC content in mapped and unmapped reads for each of the 3 kits. As shown in Table 2, no substantial difference in GC content was introduced during the preparation methods of the 3 kits.
Time Comparison
from sheared DNA to size selection
171For bulk or alternate pack sizes, email us at [email protected].
DNA Sequencing | Next Generation Sequencing Library Prep
Prep2Seq™ DNA Library Prep Kit for Illumina platform continued...
79800 1 KT Set 1(Contains adapter sequences 1-12) 2 KT Set 2(Contains adapter sequences 13-27)
Prep2Seq Multiplex Oligo Adapters allow sample multiplexing in NGS reactions. These are HPLC purified adapter duplex sets that are bar coded and designed for multiplex sample preparation for Next Generation Sequencing on Illumina platforms.
Set 1 components:12 multiplex adapters that correspond to Illumina's
adapters 1-12.20 reactions per adapter Each adapter vial contains 4.8 nmol, 80 μl.
Set 2 components:12 multiplex adapters that correspond to Illumina's
adapters 13-16, 18-23, 25, and 27. 20 reactions per adapterEach adapter vial contains 4.8 nmol, 80 μl.
Prep2Seq Multiplex Oligo Adapters for Illumina platform
TOTAL kit time
Workflow Time Comparison
from sheared DNA to size selection
Hands-on Lab Time-
Walk-Away Lab Time-
VS.
TTTTTTOOOTALTTtt tttttimimimimime
WaWW lk-Away
min Ligation set up
Illumina TruSeq Nano
NEB Next Ultra
USB Prep2Seq 5
min Ligation set up
30min Incubation
40min Incubation
60min Incubation 30min
Incubation55min
Clean up
5 min
End repair set up
30min Incubation
35 min
Clean up
5 30min Incubation
5
5 10min Incubation
90min Clean up
30min Clean up
5 min End repair/ A-tail set up
min A-tail set up
5 min End repair/ A-tail set up
210min TOTAL kit time
155min TOTAL kit time
110min TOTAL kit time
min Ligation set up
The Prep2Seq DNA Library Prep Kit ensures efficiency of ligation to T-tailed adapters by optimizing the A-tailing step. This enzymology focus on A-tailing ensures that end bias is reduced, and the formation of blunt- and G-tailed ends is minimized. Most enzymes used for A-tailing are inefficient and result in a large amount of blunt-end DNA that can ligate as concatemers, forming chimeric false joints and compromising sequencing data. Effective A-tailing is crucial in preparing efficient DNA adapter libraries that are ready for sequencing.
Also available separately are barcoded adapter sets that enable multiplexing of up to 24 samples. Prep2Seq Multiplex Oligo adapters for Illumina systems are available in kits of 1-12 and 13-27 duplex (79800 1 KT and 79800 2 KT).
Kit components:This kit is designed for input sheared genomic DNA amounts from 100 ng to 1 μg and consists of 3 tubes: Prep2Seq NGS Enzyme Mix, Prep2Seq NGS 2X Buffer, and Prep2Seq NGS High Concentration
Ligase. A streamlined workflow, straightforward pro-tocol, and high performing kit ensures quality library prep for your Illumina sequencing runs.
Per 10 reactions: Prep2Seq NGS Enzyme Mix, 40 μl Prep2Seq NGS 2X Buffer, 250 μl Prep2Seq NGS High Concentration Ligase, 40 μl
Shipping and storage:Shipped on dry ice. Store at -20°C.
WorkflowTime Comparison
from sheared DNA to size selection
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78500 100 reactions
The Thermo Sequenase Cycle Sequencing Kit allows for equally efficient incorporation of both ddNTPs and dNTPs in cycle sequencing reactions resulting in very uniform band intensities which can be read accurately. The kit offers the choice of a three-dNTP extended primer method with internal α-labeled dNTPs or a 5' end-labeled primer method (see Table 1).
Obtain high quality sequenceLike Sequenase DNA Polymerase, Thermo Sequenase generates sequences with uniform band intensities and does not have the variability in band intensities of Taq DNA polymerase(1,2).
Flexible labelingThe kit can be used for either internal labeling with α-labeled dNTPs or with 5' end-labeled primers.
Use less templateAs little as 5 fmol of single-stranded or 20 fmol of double-stranded template is needed.
Read through secondary structureRegions with palindromes, hairpins or base repeats are more easily resolved with Thermo Sequenase and cycle sequencing methods.
Resolve gel compressions with 7-deaza-dGTP nucleotide mixesWhen DNA is synthesized with 7-deaza-dGTP instead of dGTP, weaker secondary structures are formed, thus reducing compression artifacts in sequencing gels.
Eliminate weak bands with pyrophosphataseOccasional weak bands occur with prolonged reac-tion or cycle times(3,4). This is caused by sequence-specific pyrophosphorolysis which is catalyzed by the polymerase. The addition of pyrophosphatase minimizes pyrophosphorolysis and can prevent incor-rect base-calling due to weak band intensities.
Kit components:Thermo Sequenase with thermostable
pyrophosphataseControl DNA pUC18Control Primer (-40 Forward, 23-mer)
Cycle Mix 7-deaza-dGTPCycle Mix dATPCycle Mix dTTPCycle Mix dCTP
ddGTP Termination Mix (7-deaza-dGTP)ddATP Termination Mix (7-deaza-dGTP)ddTTP Termination Mix (7-deaza-dGTP)ddCTP Termination Mix (7-deaza-dGTP)
Reaction Buffer (concentrate)Stop Solution
Shipping and storage:Shipped on dry ice. Store at -20°C.
Important notes:The formulation of Thermo Sequenase DNA Polym-erase in this kit requires the use of a glycerol tolerant DNA sequencing gel to eliminate glycerol-induced distortion of bands at approximately 350 to 600 bases beyond the primer. Information on use is included in the kit protocol. If this region is beyond the reading range, Tris-Borate-EDTA Buffer (TBE) may be used.
References:1. Tabor, S. and Richardson, C. C. (1995) Proc.Natl.
Acad.Sci.USA 92, 6339-6343.2. Samols, S. B., McArdle, B. F., Ruan, C. C.,
Vanderhorn, P. B. and Fuller, C. W. (1995) Comments 22, No. 2, United States Biochemical Corp., Cleveland, OH.
3. Ruan, C. C., Samols, S. B. and Fuller, C. W. (1990) Comments 17, No.1, United States Biochemical Corp., Cleveland, OH.
4. Tabor, S. and Richardson, C. C. (1990) J.Biol.Chem. 265, 6447-6458.
Related products
ExoSAP-IT® PCR Product Cleanup78250 20 reactions78200 100 reactions78201 500 reactions
PrepEase® Quick MiniSpin Plasmid Kit78742 250 preps
RapidGel, 40% Liquid Acrylamide Stock Solution75848 500 ml
Glycerol Tolerant Gel Buffer, 20X Solution75827 1 L
TBE Buffer, 5X Solution75891 1 L 5 L
DNA Sequencing | Thermo Sequenase Kits
Thermo Sequenase Cycle Sequencing Kit
Fig. 1. Two step cycle sequencing of plasmid DNA using the Thermo Sequenase Cycle Sequencing Kit and α-33P-dATP.
G A T C
Table 1. Cycle sequencing requirements
MethodLabel useda
Sensitivityb μg (fmol) M13
Special requirements
Typical use
Total timec
3-dNTPInternal labelcycle protocol
5µCiα-dATP
≥ 0.01 μg (5 fmol)
Must design primer for cycled labeling.
Direct sequencing of smallest quantities of less-pure DNA.
3–4 hours
Radiolabeled primer cycle protocol
γ-dATP ≥ 0.01 µg (5 fmol) (33P)
T4 Polynucleotide Kinase, γ-ATP for labeling primer.
Repeated sequencing of similar templates.
2–3 hours
a In most cases, internal (α-dATP) labeling can be performed using 35S, 33P, or 32P-labeled dATP or dCTP.b Nominal minimum quantities of M13 DNA giving clear, reliable sequence with overnight exposure on film using 33P labeled nucleotide.c Including cycling time. Hands-on time is approximately 30 - 40 minutes for all methods.
173For bulk or alternate pack sizes, email us at [email protected].
DNA Sequencing | Thermo Sequenase Kits
79260 50 reactions
The Thermo Sequenase Dye Primer Manual Cycle Sequencing Kit is designed to be used with fluorescent dye-labeled primers and high-resolution fluorescence scanners. Sequencing reactions involve a termination step followed by standard slab gel electrophoresis and scanning of the gel to detect fluorescently tagged sequencing products.
Direct sequencingSequence from bacterial colonies, M13 or lambda plaques. Single yeast patches on plates or small liquid culture may be sequenced by using this kit combined with the PrepEase Yeast Plasmid Isolation Kit (PN 79220).
Single-step protocolSequencing reactions involve use of fluorescent-labeled primers and a termination step. Note: The choice of fluorescent dyes depends on the scanner. For instruments equipped with a 532 nm laser, TAMRA (Rhodamine dye), HEX (fluorescein dye), ROX, and FAM labeled primers may be used.
Flexible detectionThe gel can be scanned without opening the glass plates, thus, allowing gel electrophoresis to be stopped at any point for analysis and then resumed if longer read lengths are needed.
Thermo sequenase DNA polymerase/tap enzyme mixThe enzyme is a mixture of Thermo Sequenase and thermostable inorganic pyrophosphatase (TAP) cloned from the thermophile Thermoplasmaacidophilum. TAP prevents pyrophosphorolysis from occurring. Pyrophosphorolysis can result in faint bands, which can affect the accuracy of base calling.
Resolve gel compressions with 7-deaza-dGTP nucleotide mixesWhen DNA is synthesized with 7-deaza-dGTP instead of dGTP, weaker secondary structures are formed, thus reducing compression artifacts in sequencing gels.
Specialized formamide loading dyeA special formulation of the two tracking dyes does not obscure TAMRA or HEX primers. The faster migrating dye fluoresces at a wavelength greater than 649 nm. The slower migrating dye fluoresces at a wavelength greater than 600 nm. Thus, sequences will not be obscured if TAMRA or HEX labeled prim-ers are used.
Direct loading of samples on the gel Alcohol precipitation of the sequencing reaction products is not required. Simply load the reaction onto a standard sequencing gel.
Important notes:The formulation of Thermo Sequenase DNA Polymerase in this kit necessitates the use of a glycerol tolerant DNA sequencing gel to eliminate glycerol-induced distortion of bands at approximate-ly 350 to 600 bases beyond the primer. Information on use is included in the kit’s protocol. If this region is beyond the reading range, Tris-Borate-EDTA Buffer (TBE) may still be used.
Kit components:Thermo Sequenase with thermostable inorganic
Pyrophosphatase Reaction Buffer (concentrate) Control DNA pUC19Control Primer, TAMRA -40 Universal Forward PrimerddG Termination Mix (7-deaza-dGTP)ddA Termination Mix (7-deaza-dGTP)ddT Termination Mix (7-deaza-dGTP)ddC Termination Mix (7-deaza-dGTP)Formamide Loading Dye
Shipping and storage:Shipped on dry ice. Store at -20°C.
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ExoSAP-IT PCR Product Cleanup78250 20 reactions78200 100 reactions78201 500 reactions
Glycerol Tolerant Gel Buffer, 20X Solution75827 1 L
PrepEase BAC Purification Kit78722 10 preps
PrepEase Plasmid 8-well Strip Kit78745 12 x 8 strips
PrepEase Quick MiniSpin Plasmid Kit78742 250 preps
RapidGel, 40% Liquid Acrylamide Stock Solution75848 500 ml
TBE Buffer, 5X Solution75891 1 L 5 L
PrepEase Yeast Plasmid Isolation Kit79220 50 preps
Thermo Sequenase Dye Primer Manual Cycle Sequencing Kit
Fig. 1. Direct sequencing from single bacterial colony using the Thermo Sequenase Dye Primer Manual Cycle Sequencing Kit.
250
200
150
100
G A T C
174 888-362-2447 | 216-765-5000 | usb.affymetrix.com
DNA Sequencing | Sequenase Polymerase
70775Y 200 units70775Z 1,000 unitsCustoms by request
Sequenase Version 2.0 Reaction Buffer, 5X(Included with each pack size of enzyme)70702 1 ml
Source:E.colistrain containing clones that overproduce T7 Gene 5 Protein and E.coliThioredoxin.
Description:Sequenase Version 2.0 DNA Polymerase is a genetically engineered form of T7 DNA polymerase(1). Unlike the wild-type enzyme, it has virtually no 3'→5' exonuclease activity. Sequenase Version 2.0 is highly processive, incorporates nucleotide analogs (dlTP, thio-dNTPs, dideoxy-NTPs, etc.), is not impeded by secondary structures and can carry out strand displacement synthesis. It is an excellent enzyme for dideoxy-sequencing and is useful in other applica-tions, especially where the absence of associated exonuclease activity is desirable.
Applications:�� DNA Sequencing�� Second strand synthesis for microarray applications(2)
�� Random primed labeling with nucleotide analogs(2)
�� Labeling the 3' termini of DNA fragments with 5' protruding ends�� Amplification of DNA�� Strand displacement applications
Note: Sequenase Version 2.0 DNA Polymerase may not be useful for making blunt-end fragments because it can leave a one-base over-hang at the 3' end.
Properties:Molecular Weight: Consists of two subunits,
modified T7 gene 5 protein (76 kDa) and E.coli thioredoxin (12 kDa).
Optimum pH: 7.5Optimum Temperature: 37°CRequirements for Divalent Cation: Mg2+, Mn2+
Purity:Greater than 95% pure as determined by SDS-PAGE. Tested for contaminating double- and single-strand-ed endonucleases and exonucleases.
Storage buffer:20 mM potassium phosphate (pH 7.4), 1 mM DTT, 0.1 mM EDTA, 50% glycerol.
Assay conditions:The reaction mixture (100 µl) contains 40 mM Tris-HCl (pH 7.5), 10 mM MgCl2, 5 mM DTT, 0.3 mM dNTPs and 5 µg M13mp18 pre-annealed to 5 pmol M13 universal primer; incubation is at 37°C for 1 minute.
Unit definition:One unit of enzyme catalyzes the incorporation of 1 nmol of nucleotide into acid insoluble form in 30 seconds at 37°C.
Concentration:13 units/µl
Functional test:DNA sequencing with the Sequenase Version 2.0 DNA Sequencing Kit (PN 70770).
Functionally Tested 5X Sequenase Version 2.0 Reaction Buffer (1 ml included, PN 70702):200 mM Tris-HCl (pH 7.5), 100 mM MgCl2, 250 mM NaCl
Sequenase Version 2.0 Dilution Buffer (1 ml included):10 mM Tris-HCl (pH 7.5), 5 mM DTT, 0.1 mM EDTA
Shipping and storage:Shipped on dry ice. Store at -20°C.
References:1. Tabor, S. and Richardson, C. C. (1989) J.Biol.
Chem. 264, 6447-6458.2. Wang, D., Coscoy, L., Zylberberg, M., Avila, P. C.,
Boushey, H. A., Ganem, D. and DeRisi, J. L. (2002) Proc.Natl.Acad.Sci.USA, 99, 15687-15692.
3. Paris, M. (1992) Comments18, (No. 3), United States Biochemical Corporation, Cleveland, Ohio.
4. Tabor, S. and Richardson, C. C. (1987) Proc.Natl.AcadSci.USA84, 4767-4771.
5. Tabor, S. and Richardson, C. C. (1989) Proc.Natl.Acad.Sci.USA86, 4076-4080.
Related products
Control DNA, M13mp18, 0.2 µg/µl70704 50 µl 50 µg
DTT Solution, 0.1 M70726 150 µl
Labeling Mix for dGTP, 5X70710 100 μl
Mn Buffer72600 100 μl
Stop Solution70724 1,200 µl
ddA Termination Mix for dGTP70714 250 μl
ddA Termination Mix for dITP70728 125 µl
ddC Termination Mix for dGTP70716 250 μl
ddG Termination Mix for dGTP70718 250 μl
ddT Termination Mix for dGTP70720 250 μl
Sequenase Version 2.0 DNA Polymerase (E.C.2.7.7.7)
175For bulk or alternate pack sizes, email us at [email protected].
DNA Sequencing | Sequenase Kits
70770 100 reactions
The Sequenase Version 2.0 DNA Sequencing Kit features Sequenase Version 2.0 DNA Polymerase, the standard for high quality DNA sequencing(1). Sequenase Version 2.0 DNA polymerase is a genetically engineered form of T7 DNA polymerase which retains extraordinary polymerase activity with virtually no 3'→5' exonuclease activity. It is highly processive, able to effectively incorporate nucleotide analogs for sequencing (dideoxy NTPs, α-thio dATP, dITP, 7-deaza-dGTP, etc.) and is not easily impeded by template secondary structure. The kit includes all the reagents necessary to achieve high quality results (Fig. 1). Use of the specially formulated buf-fers and mixes included in the kit will maximize yield of sequence information.
Flexible labelingThe kit can be used for either internal labeling with α-labeled dNTPs or with 5'end-labeled primers.
Resolve gel compressions with dITP nucleotide mixesThe substitution of dITP (deoxyinosine triphosphate) for dGTP in the reaction mix eliminates the second-ary structures that produce gel compressions. dITP forms fewer H-bonds with dCTP than does dGTP, so product is more readily denatured during gel electrophoresis. Hence, sequence data is free from gel-based compression artifacts and results are more accurate.
Emphasize sequence close to primers with Mn bufferManganese (Mn) is added to emphasize sequence close to the primer, which may be weak if insuf-ficient DNA template is used. Mn++ increases the incorporation rate of dideoxynucleotides relative to deoxynucleotides. Thus, termination occurs earlier and more sequence is visible close to the primer(2,3).
Increase read length with sequence extending mixesThese mixes enable chain terminations to be extended to more than 3,000 bases from the primer. The use of these mixes provides a simple method to further extend the range of sequence, if needed. Keep in mind that this degree of extension can reach well beyond the limits of any electrophoresis gel resolution, yet use of the mixes when combined with short and long gel runs can increase overall sequence yield.
Eliminate weak bands with pyrophosphataseOccasionally, weak bands may occur with prolonged reaction times (greater than 5 minutes)(4,5), or when dITP is used in the sequencing reaction(6). The addition of pyrophosphatase can prevent weak band intensities brought on by sequence-specific pyrophosphorolysis catalyzed by the polymerase.
Convenient enzyme storage with glycerol enzyme dilution bufferSequenase DNA polymerase can be pre-diluted to a working concentration for storage of the enzyme in this form. This eliminates the necessity of diluting the polymerase prior to each sequencing reaction. Also, the addition of glycerol enhances the stability of the enzyme in sequencing reactions. Note: Pre-dilution of the polymerase with this buffer results in higher glycerol concentration in the sequencing reaction. A Glycerol Tolerant Gel (GTG) Buffer (PN 75827) must be used in the sequencing gel and buffer chambers to eliminate glycerol-induced distortion of bands at approximately 350 to 600 bases beyond the primer(7). If this region is beyond the region of inter-est, Tris-Borate-EDTA (TBE) Buffer may be used (PN 70454 or 75891).
Kit components:Sequenase Version 2.0 DNA PolymeraseInorganic Pyrophosphatase
Enzyme Dilution BufferGlycerol Enzyme Dilution BufferSequenase Reaction Buffer (5X)Dithiothreitol SolutionMn Buffer
Control DNA M13 mp18Primer (-40 Universal)
Labeling Mix (dGTP, 5X)ddGTP Termination Mix (for dGTP)ddATP Termination Mix (for dGTP)ddTTP Termination Mix (for dGTP)ddCTP Termination Mix (for dGTP)Sequencing Extending Mix (for dGTP)
Labeling Mix (dITP, 5X)ddGTP Termination Mix (for dITP)ddATP Termination Mix (for dITP)ddTTP Termination Mix (for dITP)ddCTP Termination Mix (for dITP)Sequence Extending Mix (for dITP)
Stop Solution
Shipping and storage:Shipped on dry ice. Store at -20°C.
References:1. Tabor, S. and Richardson C. C. (1989) J.Biol.
Chem. 264, 6447-6458.2. Fuller, C. W. (1989) Comments 16, No. 3, United
States Biochemical Corp., Cleveland, OH.3. Tabor, S. and Richardson, C. C. (1989) Proc.Nat.
Acad.Sci.USA 86, 4076-4080.4. Ruan, C. C., Samols, S. B. and Fuller, C. W. (1990)
Comments 17, No. 1, United States Biochemical Corp., Cleveland, OH.
5. Tabor, S. and Richardson C. C. (1990) J.Biol.Chem. 265, 8322-8328.
6. Tabor, S. and Richardson, C. C. (1989) Proc.Nat.Acad.Sci.USA 84, 4767-4771.
7. Pisa-Williamson, D. And Fuller, C. W. (1992) Comments 19, No. 2, United States Biochemical Corp., Cleveland, OH.
Sequenase Version 2.0 DNA Sequencing Kit
Fig. 1. Standard sequencing results using 35S-labeled dATP and the Sequenase Version 2.0 DNA Sequencing Kit.
176 888-362-2447 | 216-765-5000 | usb.affymetrix.com
70170 100 reactions
The Sequenase PCR Product Sequencing Kit utilizes a novel enzymatic cleanup method to pre-treat PCR products prior to sequencing without any subsequent purification or separation steps. This kit contains all of the standard Sequenase Version 2.0 Kit components with the addition of Shrimp Alkaline Phosphatase (SAP) and Exonuclease I to effectively remove excess dNTPs and primers present in the final PCR product reaction mixture. Both enzymes are added simultaneously to a specific aliquot of the PCR product mixture, incubated at 37°C and then inactivated at 80°C. The result is a cleaner PCR product, free from excessive nucleotides and primers, ready to be sequenced with standard Sequenase reagents.
Flexible labelingThe kit can be used for either internal labeling with α-labeled dNTPs or with 5'end-labeled primers.
Resolve gel compressions with 7-deaza-dGTP nucleotide mixesWhen DNA is synthesized with 7-deaza-dGTP instead of dGTP, weaker secondary structures are formed, thus reducing compression artifacts in se-quencing gels. Generally, 7-deaza-dGTP is preferred over dITP since band signal tends to be stronger and more uniform. However, dITP is still recommended for resolving the strongest gel compressions.
Eliminate weak bands with pyrophosphataseOccasionally, weak bands may occur with prolonged reaction times (greater than 5 minutes)(3,4), or when dITP is used in the sequencing reaction(5). The addition of pyrophos phatase can prevent weak band intensities brought on by sequence-specific pyrophosphorolysis catalyzed by the polymerase. The formulation of Sequenase enzyme included in this kit is pre-mixed with pyrophosphatase for added convenience.
Convenient use of sequenase enzymeSequenase Version 2.0 DNA Polymerase within this kit is pre-diluted to a working concentration. This eliminates the necessity of diluting the polymerase prior to each sequencing reaction. Also, the enzyme is stored in 50% glycerol which enhances the stabil-ity of the enzyme in the sequencing reaction.
Kit components:Sequenase Version 2.0 DNA Polymerase with
Inorganic PyrophosphataseSAP, 200 unitsExonuclease I, 1,000 units
Sequenase Reaction Buffer (5X)Dithiothreitol SolutionMn Buffer
Control DNA (M13 clone)Primer (-40 Forward 23-mer)Primer (-50 Reverse 21-mer)dGTP, 3 µM dATP, 3 µMdTTP, 3 µM dCTP, 3 µM7-deaza-dGTP, 3 µM
Labeling Mix (dGTP, 5X)ddGTP Termination Mix (for dGTP)ddATP Termination Mix (for dGTP)ddTTP Termination Mix (for dGTP)ddCTP Termination Mix (for dGTP)
Labeling Mix (7-deaza dGTP, 5X)ddGTP Termination Mix (for 7-deaza-dGTP)ddATP Termination Mix (for 7-deaza-dGTP)ddTTP Termination Mix (for 7-deaza-dGTP)ddCTP Termination Mix (for 7-deaza dGTP)
Stop Solution
Shipping and storage:Shipped on dry ice. Store at -20°C.
References:1. Fuller, C. W. (1989) Comments 16, No. 3, United
States Biochemical Corp., Cleveland, OH.2. Tabor S. and Richardson, C. C. (1989) Proc.Nat.
Acad.Sci.USA 86, 4076-4080.3. Ruan, C. C., Samols, S. B. and Fuller, C. W. (1990)
Comments 17, No. 1, United States Biochemical Corp., Cleveland, OH.
4. Tabor S. and Richardson, C. C. (1990) J.Biol.Chem. 265, 6447-6458.
5. Tabor S. and Richardson, C. C. (1987) Proc.Natl.Acad.Sci.USA 84, 4767-4771.
Sequencing kits
PCR Product Pre-Sequencing Kit70995 100 reactions70996 500 reactions70997 2,000 reactions
Sequenase Version 2.0 DNA Sequencing Kit70770 100 reactions
Sequenase Quick Denature PlasmidSequencing Kit70140 100 reactions
Thermo Sequenase Cycle Sequencing Kit 78500 100 reactions
Sequencing enzymes
Sequenase Version 2.0 DNA Polymerase 70775Y 200 units70775Z 1,000 units
PCR cleanup
ExoSAP-IT PCR Product Cleanup78250 20 reactions78200 100 reactions78201 500 reactions78202 2,000 reactions78205 5,000 reactions
HT ExoSAP-IT High-Throughput PCR Product Cleanup78395 480 reactions x 8-tube strip
5,760 reactions x 1 plate (12 x 8-tube strips)
23,040 reactions - 4 plates
1,000 reactions
5,000 reactions
Ultrapure reagents for DNA sequencing
Ammonium Persulfate76322 10 gm 100 gm 1 kg
Glycerol Tolerant Gel Buffer, 20X solution75827 1 L
RapidGel-40% Liquid Acrylamide75848 500 ml
RapidGel-XL-6%, TBE75861 500 ml
TEMED76320 100 gm 500 gm
TBE Buffer, 10X Powder70454 6 bottles
Sequenase PCR Product Sequencing Kit
DNA Sequencing | Sequenase Kits
177For bulk or alternate pack sizes, email us at [email protected].
70140 100 reactions
The Sequenase Quick Denature Plasmid Sequenc-ing Kit offers two simple and efficient denaturation methods that do not require precipitation of plasmid DNA. Both methods, Glycol/Glycerol and Alkali/Acid, allow for rapid generation of denatured template DNA suitable for use with standard Sequenase Version 2.0 DNA Polymerase. In addition, this kit contains the 7-deaza-dGTP nucleotide mixes which provide stronger band intensities for resolving sequencing gel compressions.
Rapid and simple denaturation protocolNo ethanol precipitation is needed. Simply denature plasmid template for up to 10 minutes, anneal primer for another 10 minutes and sequence.
Use less templateAs little as 0.5 µg of plasmid DNA is all that is needed to generate good sequence results.
Resolve gel compressions with 7-deaza-dGTP nucleotide mixes
Emphasize sequence close to primers with Mn bufferMn++ increases the incorporation rate of dide-oxynucleotides relative to deoxynucleotides. Thus, termination occurs earlier and more sequence is visible close to the primer(2,3).
Increase read length with sequence extending mixesThese mixes enable chain terminations to be extended to more than 3,000 bases from the primer. The use of these mixes provides a simple method to further extend the range of sequence, if needed. Keep in mind that this degree of extension can reach well beyond the limits of any electrophoresis gel resolution, yet use of the mixes when combined with short and long gel runs can increase overall sequence yield.
Eliminate weak bands with pyrophosphataseOccasionally, weak bands may occur with prolonged reaction times (greater than 5 minutes)(3,4), or when dITP is used in the sequencing reaction(5). The addition of pyrophos phatase can prevent weak band intensities brought on by sequence-specific pyrophosphorolysis catalyzed by the polymerase. The formulation of Sequenase enzyme included in this kit is pre-mixed with pyrophosphatase for added convenience.
Convenient use of sequenase enzymeSequenase Version 2.0 DNA Polymerase within this kit is pre-diluted to a working concentration. This eliminates the necessity of diluting the polymerase prior to each sequencing reaction. Also, the enzyme is stored in 50% glycerol which enhances the stabil-ity of the enzyme in the sequencing reaction.
Kit components:Sequenase Version 2.0 DNA Polymerase with
inorganic pyrophosphatasePlasmid Reaction Buffer (concentrate)Plasmid Denaturing ReagentNaOH, 1.0 M SolutionHCl, 1.0 M SolutionDithiothreitol SolutionMn Buffer
Control DNA pUC19Primer (-40 Forward, 23-mer)Primer (-50 Reverse, 21-mer)
Labeling Mix (7-deaza-dGTP, 5X)ddGTP Termination Mix (7-deaza-dGTP)ddATP Termination Mix (7-deaza-dGTP)ddTTP Termination Mix (7-deaza-dGTP)ddCTP Termination Mix (7-deaza-dGTP)
Stop Solution
Shipping and storage:Shipped on dry ice. Store at -20°C.
References:1. Fuller, C. W. (1989) Comments 16, No. 3, United
States Biochemical Corp., Cleveland, OH.2. Tabor S. and Richardson, C. C. (1989) Proc.Nat.
Acad.Sci.USA 86, 4076-4080.3. Ruan, C. C., Samols, S. B. and Fuller, C. W. (1990)
Comments 17, No. 1, United States Biochemical Corp., Cleveland, OH.
4. Tabor S. and Richardson, C. C. (1990) J.Biol.Chem. 265, 6447-6458.
5. Tabor S. and Richardson, C. C. (1987) Proc.Natl.Acad.Sci.USA 84, 4767-4771.
6. Pisa-Williamson, D. and Fuller, C. W. (1992) Comments 19, No. 2, United States Biochemical Corp., Cleveland, OH.
Sequencing kits
Sequenase Version 2.0 DNA Sequencing Kit70770 100 reactions
Sequenase PCR Product Sequencing Kit70170 100 reactions
Thermo Sequenase Cycle Sequencing Kit78500 100 reactions
Sequencing enzymes
Sequenase Version 2.0 DNA Polymerase 70775Y 200 units70775Z 1,000 units
Sequencing reagents
Glycerol Tolerant Gel Buffer, 20X solution75827 1 L
RapidGel-40%75848 500 ml
RapidGel-XL-6%75861 500 ml
Taurine, Ultrapure75824 1 kg
Sequenase Quick Denature Plasmid DNA Sequencing Kit
DNA Sequencing | Sequenase Kits
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DNA Sequencing | Sequenase Primer/Sequencing Tips
Primer for DNA Sequencing
T3 Promoter Primer
Prod. No. Product Description Primer Sequence [conc] Size
71200 T3 Promoter 5' ATTAACCCTCACTAAAG 3' 5 pmol/µl 250 pmol
Determining Primer Concentration
Approximate extinction coefficients and molecular weights for oligonucleotides
DNA Sequencing Gel Formulations
Standard sequencing gels Acrylamide/ Urea 20X Glycerol 10X TBE Gel Conc. Bis-Acrylamide (7.0 – 8.3 M) Tolerant Gel Buffer Buffer H2O
6% 5.7 gm/0.3 gm 42 – 50 gm 5 ml** - ~ 45 ml
8% 7.6 gm/0.4 gm 42 – 50 gm 5 ml** - ~ 45 ml
6% 5.7 gm/0.3 gm 42 – 50 gm - 10 ml ~ 40 ml
8% 7.6 gm/0.4 gm 42 – 50 gm - 10 ml ~ 40 ml
Dissolve, adjust volume to 100 ml with H2O, filter and de-gas. When ready to pour, add 1 ml of 10% ammonium persulfate and 25 µl TEMED (N, N, N', N' tetrameth-ylethylene-diamine).
** Use 4 ml for faster gel migration.
Formamide sequencing gels Acrylamide/ Urea 20X Glycerol 10X TBE Gel Conc. Bis-Acrylamide (7 M) Tolerant Gel Buffer Buffer Formamide H2O
6% 5.7 gm/0.3 gm 42 gm 5 ml - 30 – 40 ml ~ 15 ml
8% 7.6 gm/0.4 gm 42 gm 5 ml - 30 – 40 ml ~ 15 ml
6% 5.7 gm/0.3 gm 42 gm - 10 ml 30 – 40 ml ~ 15 ml
8% 7.6 gm/0.4 gm 42 gm - 10 ml 30 – 40 ml ~ 15 ml
Warming to 35 - 40°C may be required to dissolve completely. Adjust volume to 100 ml with H2O, filter and de-gas. When ready to pour, add 1 ml of 10% am-monium persulfate and 100 - 150 µl TEMED. These formamide-containing gels may be used for very strong compressions not resolved by 7-deaza-dGTP. They will require higher running voltage and run slower than urea-only gels. Prior to drying, these gels should be soaked in 5% acetic acid, 20% methanol to prevent swelling.
N = # of bases
ε260 ≅ 10,000 x N(M-1 cm-1)
Molecular Weight ≅ 330 x N
(OD260 ÷ ε260) x 106 = Concentration (µM)
Concentration (µM) x Molecular Weight = Concentration (ng/ml)
Example for a 17 mer: ε260 ≅ 170,000 (M-1 cm-1) Molecular Weight ≅ 5,600
A solution with OD260 = 1.0 will be: (1.0 ÷ 170,000) x 106 = 5.9 µM 5.9 x 5,600 = 33,000 ng/ml
179For bulk or alternate pack sizes, email us at [email protected].
DNA Sequencing | Sequencing Tips
DNA Sequencing Gel Formulations continued...
General guidelines for running polyacrylamide sequencing gels(Based on 100 ml gel solutions for regular, wedge and formamide gels) 40% Regular Wedge Formamide
Acrylamide:Bis-Acrylamide 19:1 19:1 19:1
10% Ammonium Persulfate 1 ml 1 ml 1 ml
TEMED 30 µl 30 µl 150 µl
Gel Running 55 Watts 55 Watts 55 Watts
Conditions 12 – 1600V 12 – 1600V 19 – 2500V
(Constant Power) ~40 mA ~40 mA ~25 mA
Secondary structure in DNA templates is a common cause of BAFLs (Bands Across Four Lanes observed in sequencing gels), also referred to as Stops. When Sequenase Version 2.0 Polymerase encounters a stable secondary structure in the template, it may dissociate from the template without incorporating a dideoxynucleotide. The fragments generated will co-migrate and cause artifact bands.
BAFLs can be eliminated by performing a chase step with TdT to remove artificially terminated fragments. Terminal Deoxynucleotidyl Transferase is a polymerase which non-specifically adds nucleotides to a free 3' hydroxyl. Its action in this case is to extend the fragments which are not properly terminated with a dideoxynucleotide. TdT will add hundreds of nucleotides in a relatively short period of time allowing non-specifically terminated fragments to be extended such that they run at the top of the gel without interfering with readable sequence.
This method allows for the complete resolution of BAFLs and difficult gel compressions.
Method1. A 1 mM solution of all four dNTPs is made in Sequenase Enzyme Dilution
Buffer. 1 µl each dNTP (100 mM stock) 96 µl Enzyme Dilution Buffer2. Terminal Deoxynucleotidyl Transferase (TdT) [PN 72033] is diluted in the
1 mM dNTP solution (above) to 2 units/µl. Each termination reaction will require 1 µl of this enzyme/dNTP dilution. Mix well on ice. It is best to prepare this enzyme/dNTP solution immediately prior to use.
3. Sequence according to the Sequenase Version 2.0 Sequencing Protocol using the appropriate labelling and termination mixes. DO NOT add Stop Solution.
4. After the 2-5 minute termination reaction is complete add 1 µl of the TdT/dNTP solution to each termination reaction tube.
5. Mix and incubate 15-30 minutes at 37°C.6. Add 4 µl of Stop Solution to each reaction tube.7. Heat denature and load on a sequencing gel as usual.
References:1. Tabor, S. and Richardson, C. C. (1987) Proc.Natl.Acad.Sci USA 84, 4767-
4771.2. Fawcett, T. W. and Bartlett, S. G. (1991) Comments, (United States
Biochemical Corporation), 17, (4) pg. 19.3. Fawcett, T. W. and Bartlett, S. G. (1990) BioTechniques 9, 46-49.
Related products
dNTP 100 mM Solutions, Set of Four4 x 25 µmol (0.25 ml) each dNTP per pack77100 1 pack
Sequenase Version 2.0 DNA Sequencing Kit70770 100 reactions
Terminal Deoxynucleotidyl Transferase, Recombinant (rTdT)72033 500 units 2,500 units
Thermo Sequenase Cycle Sequencing Kit78500 100 reactions
Use of Terminal Deoxynucleotidyl Transferase (TdT) to Resolve Bands Across Four Lanes (BAFLs)
Fig. 1. Use of TdT to eliminate BAFLs. Lane A: Plasmid sequence with 2 strong BAFLs indicated by arrows. Lane B: Sequence results with same plasmid that included a TdT/dNTP chase reaction.
A B
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DNA Sequencing | Sequencing Tips
Compression artifacts may be produced during electrophoresis due to the formation of stable secondary structures in the sequencing product DNA fragments. The folded structures run faster through the gel matrix than equivalent unfolded DNA fragments; consequently they will migrate to the same position as smaller sized fragments resulting in bands across 2 or more lanes. Typical indications of a compression are odd spacing, bands very close together followed by bands spaced farther apart than normal, or bands in two to four lanes at the same point. Sometimes bases can be lost entirely. The most difficult instances are those in which the compressed area looks perfectly normal. These are detected when the opposite strand is sequenced or nucleotide analogs are used to check for compression artifacts. Nucleotide analogues such as 7-deaza-dGTP or dITP can be used to alleviate compressions but in some instances are not able to fully resolve a difficult compression. In these cases, the use of a formamide gel alone or in conjunction with the nucleotide analogue, will completely relieve the compressed areas giving unambiguous sequence.
MethodPreparation and running a formamide gel is not very different from a standard sequencing gel. A 6% or 8% acrylamide gel may be prepared. The running time is increased to approximately one and one-half times that of a standard gel.
1. Weigh the following dry ingredients into a beaker: 6% gel 5.7 gm Acrylamide 0.3 gm N-N' Methylene-bis-acrylamide 42 gm Urea
8% gel 7.6 gm Acrylamide 0.4 gm N-N' Methylene-bis-acrylamide 42 gm Urea
2. Add: 40 ml Formamide 10 ml 10X TBE Buffer OR 5 ml 20X GTG Buffer (for a Glycerol Tolerant Gel)
and a magnetic stir bar
3. Warm the gel mix to 50°C and stir until the components are completely dissolved.
4. Continue stirring until the temperature cools to ~30°C. Adjust the volume to 100 ml with distilled water and vacuum filter with a nitrocellulose filter unit. The gel should be poured immediately. To prevent the urea from precipitating, do not allow the gel to cool below 25°C. If the mix should happen to precipitate the process must be repeated from the beginning; however running a gel that has been redissolved will give very poor results.
5. Add 1 ml of 10% ammonium persulfate and 25 µl of TEMED. (NOTE: Use this amount of TEMED when pouring the gel the night before it is to be run. If the gel is to be used the same day, adding four to five times the amount of TEMED will polymerize the gel in an hour. It is usually more convenient to pour the gel at the end of one day and run it first thing the next day to accommodate the longer running time of formamide gels). Mix well and pour immediately into clean gel plates. The solution is quite viscous and somewhat more difficult to pour than a standard gel. It may be necessary to tip the plates completely upright to get the gel mix to spread properly between them.
6. When preparing the gel for electrophoresis, be sure to clean out the wells thoroughly prior to loading the samples. Run the gel at 60W constant power. Expect the voltage to climb to 2,000V and the plates to reach a higher temperature than usual as the gel progresses. Normally the dye fronts eventually run as thin bands.
7. Stop the gel at the desired point and soak it in a 10% acetic acid, 20% methanol solution for 45 minutes or until shrinkage is apparent. Formamide gels tend to swell when placed into the soaking solution, however, with time they will shrink to a manageable size. Dry and expose the gel as you would any other. In some cases the formamide gel may require slightly longer drying time than a standard gel.
Relief from Compression Artifacts in DNA Sequencing Using Formamide Gels
Fig. 1. Sequence determination using Sequenase Version 2.0 DNA Polymerase of the single-stranded vector M13mp18. A primer at position 6511 is used to visualize a very signifi-cant gel compression. A. Sequence determined using dGTP and 7-deaza-dGTP nucleotide mixes and run on a 6% TBE gel. Note the significant compression in both reactions. The sequence is unreadable at this point. B. The same reactions run on an 8% TBE/40% Formamide gel. Note the G doublet is not completely resolved in the dGTP reaction even on this Formamide gel while the 7-deaza-dGTP reaction is completely resolved.
A B
7-deaza dGTP dGTP dGTP
7-deaza dGTP
Related products
Acrylamide75820 100 gm 500 gm 1 kg
Ammonium Persulfate (APS)76322 10 gm 100 gm
bis-Acrylamide75821 25 gm 50 gm 100 gm
Formamide75828 500 gm 1 kg
TEMED76320 100 ml
Urea75826 1 kg 5 kg
181For bulk or alternate pack sizes, email us at [email protected].
DNA Sequencing | Sequencing Tips
Prevent Weak Sequencing Band Intensities by Using Pyrophosphatase
The determination of DNA sequence using a DNA polymerase (Klenow) and nucleotide-specific chain-termination was first described by F. Sanger etal.(1). In this procedure, DNA synthesis is initiated at a unique priming site, and terminated by the incorporation of 2',3'-dideoxynucleoside monophosphates (ddNMPs). Tabor and Richardson(2) noted that under certain conditions, the intensities of some of the bands on resulting sequencing gels can be weak. This phenomenon is particularly apparent when the reactions are run for 30 minutes or longer and when dITP is used in place of dGTP. These weak bands are the result of pyrophosphorolysis(3,4). This is the reversal of the polymerization reaction catalyzed by the DNA polymerase (see below). Pyrophosphorolysis can be prevented using inorganic pyrophosphatase to hydrolyze the pyrophosphate(3,4).
Reactions of DNA PolymerasesAll known DNA polymerases catalyze the template-dependent incorporation of a deoxynucleotide onto the 3' hydroxyl terminus of a primer, with the re-lease of inorganic pyrophosphate (PPi). Catalysis requires the presence of a divalent cation, either Mg2+ or Mn2+. Under the conditions normally used for DNA synthesis (i.e., high dNTP concentrations and low pyrophosphate con-centration), the forward reaction (polymerization) is greatly favored over the reverse reaction (pyrophosphorolysis). Many DNA polymerases also catalyze the hydrolysis of nucleotides from the 3' terminus of DNA through a sepa-rate exonuclease activity. This reaction is unlike pyrophosphorolysis because the substrate is H2O and the product is a nucleoside monophosphate.
The polymerization, pyrophosphorolysis and exonuclease reactions are sum-marized below:
M2+
Polymerization: DNA(n) + dNTP → DNA(n+1) + PPi
M2+
Pyrophosphorolysis: DNA(n) + dNTP ← DNA(n+1) + PPi
M2+
Exonuclease Reaction: DNA(n) + H2O → DNA(n-1) + dNMP (M2+ = Mg2+ or Mn2+)
Although pyrophosphorolysis is much slower than polymerization, it can affect DNA sequencing reactions(3,4). Some bands become less intense when the termination reactions are allowed to incubate for extended periods of time. When the reactions are incubated for 2 minutes, the bands have uni-form intensities; however, after a 30 minutes incubation, approximately one in ten bands is reduced in intensity. Note that the rates at which individual bands decrease in intensity varies widely, depending on the DNA sequence.
Sequence-specific decreases in band intensity occur even when sequenc-ing with DNA polymerases that have no exonuclease activity such as AMV Reverse Transcriptase and Sequenase Version 2.0 DNA Polymerase(5). Band weakening is stimulated by the addition of pyrophosphate to the sequenc-ing reactions and it is prevented by the addition of inorganic pyrophos-phatase or high concentrations of dNTPs(3,4). Thus, the decrease in intensity of some bands with longer incubation times is due to pyrophosphorolysis.
Action of Inorganic PyrophosphataseInorganic pyrophosphatase catalyzes the hydrolysis of pyrophosphate to two molecules of orthophosphate.
Mg2+
P2O74- + H2O → 2 HPO4
2-
This enzyme plays an important role in DNA synthesis invivo, rendering DNA polymerization essentially irreversible by removing pyrophosphate. The addition of inorganic pyrophosphatase to DNA sequencing reactions com-pletely stabilizes all band intensities, even when incubating the reactions for 60 minutes and when dITP is used in place of dGTP.
For convenience, the USB Sequenase Version 2.0 DNA Sequencing Kit (PN 70770) includes nuclease-free yeast inorganic pyrophosphatase. Pyrophosphatase can be used for sequencing with dGTP or its analogs (dITP, 7-deaza-dGTP, etc.) and is effective in the presence of Mg2+ or Mn2+. The use of both Sequenase polymerase and pyrophosphatase assures the stability of all DNA generated in the DNA sequencing reaction, providing additional assurance that the sequence determined is accurate.
References:1. Sanger, F., Nicklen, S. and Coulson, A. R. (1977) Proc.Natl.Acad.Sci.USA
74, 5463-5467.2. Tabor, S. and Richardson, C. C. (1987) Proc.Nat.Acad.Sci.Usa 84, 4767-
4771.3. Tabor, S. and Richardson, C. C. (1990) J.Biol.Chem. 265, 8322-8328.4. Ruan, C. C. Samols, S. B. and Fuller, C. W. (1990) Comments17, No. 1,
1-27.5. Tabor, S. and Richardson, C. C. (1989) J.Biol. Chem. 264, 6447-6458.
Related products
Sequenase Version 2.0 DNA Sequencing Kit70770 100 reactions
BiochemicalsAcrylamides
ACS Biochemicals
Agaroses
Albumins
Amino Acids
Antibiotics
Bacterial Culture Media
Buffers
Enzymes
Proteins
Substrates
Sugars
Ultrapures
Vitamins
Biochemicals
183For bulk or alternate pack sizes, email us at [email protected].
Formulas and formula weights are provided for compounds having a specific known structure. The values used to calculate formula weights are taken from the 2001 Table of Atomic Weights, (2003) Pure Appl. Chem., 75, 1107-1122.
To simplify grade selection for customers the following grade definitions are provided. We have selected grade names which are self explanatory, as well as those which reflect various industry standards. If any changes are noticed in a familiar grade designation for a given chemical, it is a result of a change to the grade definition rather than a change in the quality of the chemical.
USB Ultrapures are specially selected biochemicals and reagents of high quality, suitable for use in numerous biotechnology applications. These re-agents meet stringent requirements, such as adherence to compendial refer-ences where applicable, have ultra-low levels of trace metal contamination, and low levels of DNase, RNase, protease, and endotoxins where applicable.
Reference constants for nucleotides and nucleosidesValues for reference constants are taken from the literature. They are useful in determining the concentration of stock solutions prepared in the laboratory.
DBR: “Data for Biochemical Research” Dawson, R. M. C., Elliot, D. C., Elliot, W. H. , Jones, K. M., (1986) Oxford Science Publications.
NRC: “Specifications and Criteria for Biochemical Compounds,” (1972) National Academy of Sciences, Washington, D.C. 3rd edition.
Biochemical grade definitionsACS Reagent Grade: Meets the limits of purity for inorganic chemicals as established by the American Chemical Society.
CP Grade (Chemically Pure): Meets the requirements of the Food Chemicals Codex, but not intended for food use.
Genetic Performance Certified (GPC): Functionally tested in specific molecular biology applications (e.g. nucleic acid recovery, in-gel restriction enzyme digestion or ligation) to ensure optimal agarose performance.
Hi-Res™: Agaroses especially selected for high resolution of samples.
USBioAnalyzed: Meets USP or NF chemical testing requirements but is not sold as USP or NF Grade; USP and NF Grades (United States Pharmacopeia and National Formulary) must meet the requirements of those compendia and will fall under this category.
USP Grade: Meets the requirements of the U.S. Pharmacopeia.
NF Grade: Meets the requirements of the National Formulary.
MB Grade (Molecular Biology): Applies to products that are suitable for a variety of Molecular Biology applications including those which require ultra-low levels of trace metal contamination, and low levels of DNase, RNase, and protease. These products are also functionally tested when applicable.
Reagent Grade: Suitable for use in general laboratory applications; meets in-house established limits in the absence of any compendial reference for the given compound. In-house established limits and their associated test methods are in many cases derived from common compendial methods such as USP, NF, ACS, etc.
High Purity Grade: Generally meets more stringent requirements than Reagent Grade products, such as higher assay limits or lower levels of trace contaminants. When available, this grade is more suitable as a reference material than the Reagent Grade.
A Few Notes About USB Biochemicals
USB® has been supplying bulk biochemicals to diagnostic, biotech, and pharmaceutical companies for over 40 years.
�� Bulk quantities available�� Lot reserve and multiple lot testing�� Long term supply agreements�� MSDS and lot-specific Certificates of Analysis (C of As)�� Scale from small to large production runs�� Complete QA and QC assays
Consistency and superior quality are essential. Turn to USB as your single sourcing solution for raw materials, and to ensure your supply meets demand.
To learn more, visit usb.affymetrix.com/bulk.
Bulk Biochemicals
Biochemicals
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Ac-Co A ............Acetyl-coenzyme A iCDH ................... Isocitrate dehydrogenaseAc-P .................Acetyl phosphate IDP ...................... Inosine-5'-diphosphateAChE ................Acetylcholinesterase IMP ..................... Inosine-5'-monophosphateADH .................Alcohol dehydrogenase INT ...................... Iodonitrotetrazolium violetADP ..................Adenosine-5'-diphosphate ITP ...................... Inosine-5'-triphosphateALD ..................Fructose-1, 6-diphosphate aldolase dITP .................... 2'-Deoxyinosine-5'-triphosphateAIDH.................Aldehyde dehydrogenase ddITP .................. 2',3'-Dideoxyinosine-5'-triphosphateAMP .................Adenonsine-5'-monophosphate LDH ..................... L-Lactate dehydrogenaseD-AOD ..............D-Amino acid oxidase D-LDH ................. D-Lactate dehydrogenaseL-AOD ..............L-Amino acid oxidase MDH ................... L-Malate dehydrogenaseAP ....................Alkaline phosphatase MK ...................... MyokinaseASOD ...............Ascorbate oxidase MPI ..................... Mannosephosphate isomeraseATP ...................Adenosine-5'-triphosphate MUTARO ............. MutarotasedATP .................2'-Deoxyadenosine-5'-triphosphate NAD+ ................. Nicotinamide-adenine dinucleotideddATP ...............2',3'-Dideoxyadenosine-5'-triphosphate NADH ................. Nicotinamide-adenine dinucleotide, reducedATPase ..............Adenosine-5'-triphosphate NADP+ ............... Nicotinamide-adenine dinucleotide phosphateBAEE ................Benzoyl-L-arginine ethyl ester NADPH ............... Nicotinamide-adenine dinucleotide phosphate,BAPNA .............N—Benzoyl-L-arginine-p-nitroanilide reducedt-Boc ................tert-butoxy carbonyl NaOAc ................ Sodium acetateBSA ..................Bovine serum albumin NBT..................... Nitro-Blue-tetrazolium saltCBZ ..................Carbonbenzoxy NPS ..................... Nitrophenyl sulphenylChE ..................Cholinesterase 2-OG(α-KG) ........ 2-OxoglutarateCK(CPK) ............Creatine kinase (α-Ketoglutarate)CoA(CoA-SH) ....Coenzyme A PALP ................... Pyridoxal-5-phosphateCP ....................Creatine phosphate PAMP .................. Pyridoxamine-5-phosphateCTP ..................Cytidine-5'-triphosphate PC ....................... Paper chromatographydCTP.................2'-Deoxycytidine-5'-triphosphate PEP ..................... Phosphoenol pyruvateddCTP...............2'-3'-Dideoxycytidine-5'-triphosphate 6-PGDH .............. 6-Phosphogluconate dehydrogenaseDAP ..................Dihydroxyacetone phosphate 6-PG ................... 6- Phosphogluconate,DNA .................Deoxyribonucleic acid (D-Gluconate-6-phosphate)DNase ..............Deoxyribonuclease PGI ...................... Phosphoglucose isomerase,DNP ..................Dinitrophenyl (Phosphohexose isomerase),DNPyr ...............Dinitropyridyl (Glucosephosphate isomerase)DTE ..................Dithioerythritol PGK .................... 3-Phosphoglycerate kinaseDTT ...................Dithiothreitol PGIuM ................ PhosphoglucomutaseEDTA ................Ethylenediaminetetraacetic acid PGM ................... Phosphoglycerate mutaseENOL ................Enolase Pi ........................ Inorganic phosphateFAD ..................Flavin-adenine dinucleotide PK ....................... Pyruvate kinaseFDH ..................Formate dehydrogenase PL-A .................... Phospholipase AFDP ..................D-Fructose-1,6-diphosphate PL-D .................... Phospholipase DFDPase .............Fructose diphosphatase POD .................... PeroxidaseFMN .................Flavin mononucleotide Ppase .................. Pyrophosphatase, inorganicF-1-P ................D-Fructose-1-phosphate Ppi ...................... Inorganic pyrophosphateF-6-P ................D-Fructose-6-phosphate PPO..................... 2,5-Diphenyl oxazoleF-6-PK ..............Fructose-6-phosphate kinase PTA ..................... PhosphotransacetylaseFUM .................Fumarase RNA .................... Ribonucleic acidGal-1-P .............Galactose-1-phosphate tRNA ................... Transfer ribonucleic acidGAP ..................Glyceraldehyde-3-phosphate RNase ................. RibonucleaseGAPDH .............D-Glyceraldehyde-3-phosphate dehydrogenase SDH .................... Sorbitol dehydrogenase,α-GDH .............α-Glycerol phosphate dehydrogenase, (Polyol dehydrogenase) (Glycerol-1-phosphate dehydrogenase), SP ....................... Sucrose phosphorylase (Glycerol-3-phosphate dehydrogenase) TALD ................... TransaldolaseGDP ..................Guanosine-5'-diphosphate TIM ..................... Triosephosphate isomeraseGK ....................Glycerokinase Tris ...................... Tris-hydroxymethyl-aminomethaneGIDH ................L-Glutamate dehydrogenase TTP (dTTP) ........... Thymidine-5'-triphosphateGMP .................Guanosine-5'-monophosphate (2'-Deoxythymidine-5'-triphosphate)GOD .................Glucose oxidase ddTTP ................. 2',3'-dideoxythymide-5'-triphosphateGOT ..................Glucamate-oxaloacetate transaminase UDP .................... Uridine-5'-diphosphateG-1-P................D-Glucose-1-phosphate UDPG .................. Uridine-5'-diphosphate glucoseG-1,6-P
2 ...........D-Glucose-1,6-diphosphate UDPgal................ Uridine-5'-diphosphate galactose
G-6-P................D-Glucose-6-phosphate UDPGP ................ Uridine-5'-diphosphate glucose pyrophosphorylaseG-6-PDH ...........Glucose-6-phosphate dehydrogenase (Glucose-1-phosphate uridylyltransferase)GPT ..................Glutamate pyruvate transaminase UMP ................... Uridine-5'-monophosphateGR ....................Glutathione reductase UTP ..................... Uridine-5'-triphosphateGSH ..................Glutathione, reduced dUTP ................... 2'-Deoxyuridine-5'-triphosphateGSSG ................Glutathione, oxidized ddUTP ................. 2',3'-Dideoxyuridine-5'-triphosphateγ-GT .................γ-Glutamyltransferase XOD .................... Xanthine oxidaseGTP ..................Guanosine-5'-triphosphate dGTP ................2'-Deoxyguanosine-5'-triphosphate DBR: “Data For Biochemical Research” Dawson, R. M. C., ddGTP ..............2',3'-Dideoxyguanosine-5'-triphosphate Elliot, D. C.,Elliot, W. H., Jones, K. M., Oxford Science Publications (1986) HK ....................Hexokinase NRC: “Specifications and Criteria for Biochemical Compounds.” National HPLC ................High performance liquid chromatography Academy of Sciences, Washington, D.C., 3rd edition (1972).
Biochemical Acronyms
Biochemicals
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Standard Atomic Weights of the Elements
Atomic Number Element Symbol Weight
89. . . . . . . . . Actinium . . . . . . . . .Ac . . . . . . . . .227.0277 13. . . . . . . . . Aluminum. . . . . . . . . Al . . . . . . . . . . 26.981539 95. . . . . . . . . Americium . . . . . . . .Am . . . . . . . . .243.0614* 51. . . . . . . . . Antimony . . . . . . . . .Sb . . . . . . . . . .121.75 18. . . . . . . . . Argon . . . . . . . . . . . . Ar . . . . . . . . . . 39.948 33. . . . . . . . . Arsenic . . . . . . . . . . . As. . . . . . . . . . . 74.92159 85. . . . . . . . . Astatine . . . . . . . . . .At . . . . . . . . . .209.9871* 56. . . . . . . . . Barium . . . . . . . . . . .Ba . . . . . . . . . .137.327 97. . . . . . . . . Berkelium . . . . . . . . .Bk . . . . . . . . . .247.0703* 4. . . . . . . . . . Beryllium . . . . . . . . .Be . . . . . . . . . . . . 9.012182 83. . . . . . . . . Bismuth . . . . . . . . . .Bi . . . . . . . . . .208.98037 107 . . . . . . . Bohrium . . . . . . . . . .Bh. . . . . . . . . .264.12 5. . . . . . . . . . Boron . . . . . . . . . . . .B. . . . . . . . . . . . 10.811 35. . . . . . . . . Bromine . . . . . . . . . .Br . . . . . . . . . . . 79.904 48. . . . . . . . . Cadmium . . . . . . . . .Cd. . . . . . . . . .112.411 55. . . . . . . . . Cesium . . . . . . . . . . .Cs . . . . . . . . . .132.90543 20. . . . . . . . . Calcium . . . . . . . . . .Ca. . . . . . . . . . . 40.078 98. . . . . . . . . Californium. . . . . . . .Cf . . . . . . . . . .251.0796* 6. . . . . . . . . . Carbon . . . . . . . . . . .C. . . . . . . . . . . . 12.011 58. . . . . . . . . Cerium . . . . . . . . . . .Ce. . . . . . . . . .140.115 17. . . . . . . . . Chlorine . . . . . . . . . .Cl . . . . . . . . . . . 35.4527 24. . . . . . . . . Chromium. . . . . . . . .Cr . . . . . . . . . . . 51.9961 27. . . . . . . . . Cobalt . . . . . . . . . . .Co. . . . . . . . . . . 58.9332 29. . . . . . . . . Copper . . . . . . . . . . .Cu. . . . . . . . . . . 63.546 96. . . . . . . . . Curium . . . . . . . . . . .Cm . . . . . . . . .247.0704* 105 . . . . . . . Dubnium. . . . . . . . . .Db. . . . . . . . . .262.1141 66. . . . . . . . . Dysprosium. . . . . . . .Dy . . . . . . . . . .162.5 99. . . . . . . . . Einsteinium . . . . . . . .Es . . . . . . . . . .252.0830 68. . . . . . . . . Erbium . . . . . . . . . . .Er . . . . . . . . . .167.26 63. . . . . . . . . Europium . . . . . . . . .Eu . . . . . . . . . .151.965 100 . . . . . . . Fermium . . . . . . . . . .Fm . . . . . . . . .257.0951 9. . . . . . . . . . Fluorine . . . . . . . . . .F . . . . . . . . . . . . 18.9984032 87. . . . . . . . . Francium. . . . . . . . . .Fr . . . . . . . . . .223.0197 64. . . . . . . . . Gadolinium. . . . . . . .Gd. . . . . . . . . .157.25 31. . . . . . . . . Gallium . . . . . . . . . .Ga. . . . . . . . . . . 69.723 32. . . . . . . . . Germanium. . . . . . . .Ge. . . . . . . . . . . 72.61 79. . . . . . . . . Gold . . . . . . . . . . . . .Au. . . . . . . . . .196.96654 72. . . . . . . . . Hafnium . . . . . . . . . .Hf . . . . . . . . . .178.49 108 . . . . . . . Hassium . . . . . . . . . .Hs 2. . . . . . . . . . Helium . . . . . . . . . . .He. . . . . . . . . . . . 4.002602 67. . . . . . . . . Holmium . . . . . . . . .Ho. . . . . . . . . .164.93032 1. . . . . . . . . . Hydrogen . . . . . . . . .H. . . . . . . . . . . . . 1.00794 49. . . . . . . . . Indium . . . . . . . . . . . In . . . . . . . . . .114.82 53. . . . . . . . . Iodine . . . . . . . . . . . . I . . . . . . . . . . .126.90447 77. . . . . . . . . Iridium . . . . . . . . . . . Ir . . . . . . . . . . .192.22 26. . . . . . . . . Iron . . . . . . . . . . . . .Fe . . . . . . . . . . . 55.847 36. . . . . . . . . Krypton . . . . . . . . . .Kr . . . . . . . . . . . 83.8 57. . . . . . . . . Lanthanum . . . . . . . .La . . . . . . . . . .138.9055 103 . . . . . . . Lawrencium . . . . . . .Lr . . . . . . . . . .262.1097 82. . . . . . . . . Lead . . . . . . . . . . . . .Pb . . . . . . . . . .207.2 3. . . . . . . . . . Lithium . . . . . . . . . . .Li . . . . . . . . . . . . 6.941 71. . . . . . . . . Lutetium . . . . . . . . . .Lu . . . . . . . . . .174.967 12. . . . . . . . . Magnesium. . . . . . . .Mg . . . . . . . . . . 24.305 25. . . . . . . . . Manganese. . . . . . . .Mn . . . . . . . . . . 54.93805 109 . . . . . . . Meitnerium . . . . . . . .Mt. . . . . . . . . .268.1388 101 . . . . . . . Mendelevium . . . . . .Md . . . . . . . . .258.0984* 80. . . . . . . . . Mercury . . . . . . . . . .Hg. . . . . . . . . .200.59 42. . . . . . . . . Molybdenum. . . . . . .Mo . . . . . . . . . . 95.94 60. . . . . . . . . Neodymium . . . . . . .Nd . . . . . . . . .144.24
Atomic Number Element Symbol Weight
10. . . . . . . . . Neon. . . . . . . . . . . . .Ne. . . . . . . . . . . 20.1797 93. . . . . . . . . Neptunium . . . . . . . .Np. . . . . . . . . .237.0482* 28. . . . . . . . . Nickel . . . . . . . . . . . .Ni . . . . . . . . . . . 58.69 41. . . . . . . . . Niobium . . . . . . . . . .Nb. . . . . . . . . . . 92.90638 7. . . . . . . . . . Nitrogen . . . . . . . . . .N. . . . . . . . . . . . 14.00674 102 . . . . . . . Nobelium . . . . . . . . .No. . . . . . . . . .259.1010 76. . . . . . . . . Osmium . . . . . . . . . .Os . . . . . . . . . .190.2 8. . . . . . . . . . Oxygen . . . . . . . . . . .O. . . . . . . . . . . . 15.9994 46. . . . . . . . . Palladium . . . . . . . . .Pd . . . . . . . . . .106.42 15. . . . . . . . . Phosphorus . . . . . . .P . . . . . . . . . . . . 30.973762 78. . . . . . . . . Platinum . . . . . . . . . .Pt . . . . . . . . . .195.08 94. . . . . . . . . Plutonium . . . . . . . . .Pu . . . . . . . . . .244.0642* 84. . . . . . . . . Polonium. . . . . . . . . .Po . . . . . . . . .208.9824* 19. . . . . . . . . Potassium . . . . . . . . .K . . . . . . . . . . . 39.0983 59. . . . . . . . . Praseodymium . . . . .Pr . . . . . . . . . .140.90765 61. . . . . . . . . Promethium . . . . . . .Pm . . . . . . . . .144.9127* 91. . . . . . . . . Protactinium . . . . . . .Pa . . . . . . . . . .231.0359 88. . . . . . . . . Radium . . . . . . . . . .Ra . . . . . . . . . .226.0254* 86. . . . . . . . . Radon. . . . . . . . . . . .Rn. . . . . . . . . .222.0176* 75. . . . . . . . . Rhenium . . . . . . . . . . Re . . . . . . . . .186.207 45. . . . . . . . . Rhodium. . . . . . . . . .Rh. . . . . . . . . .102.9055 37. . . . . . . . . Rubidium . . . . . . . . .Rb. . . . . . . . . . . 85.4678 44. . . . . . . . . Ruthenium . . . . . . . .Ru. . . . . . . . . .101.07 104 . . . . . . . Rutherfordium. . . . . .Rf . . . . . . . . . .261.1088 62. . . . . . . . . Samarium . . . . . . . . .Sm . . . . . . . . .150.36 21. . . . . . . . . Scandium . . . . . . . . .Sc . . . . . . . . . . . 44.95591 106 . . . . . . . Seaborgium. . . . . . . .Sg . . . . . . . . . .266.1219 34. . . . . . . . . Selenium. . . . . . . . . .Se . . . . . . . . . . . 78.96 14. . . . . . . . . Silicon. . . . . . . . . . . .Si . . . . . . . . . . . 28.0855 47. . . . . . . . . Silver. . . . . . . . . . . . .Ag. . . . . . . . . .107.8682 11. . . . . . . . . Sodium . . . . . . . . . . .Na. . . . . . . . . . . 22.989768 38. . . . . . . . . Strontium . . . . . . . . .Sr . . . . . . . . . . . 87.62 16. . . . . . . . . Sulfur . . . . . . . . . . . .S . . . . . . . . . . . 32.066 73. . . . . . . . . Tantalum. . . . . . . . . .Ta . . . . . . . . . .180.9479 43. . . . . . . . . Technetium . . . . . . . .Tc . . . . . . . . . . . 97.9072* 52. . . . . . . . . Tellurium. . . . . . . . . .Te . . . . . . . . . .127.6 65. . . . . . . . . Terbium. . . . . . . . . . .Tb . . . . . . . . . .158.92534 81. . . . . . . . . Thallium . . . . . . . . . .Tl . . . . . . . . . .204.3833 90. . . . . . . . . Thorium . . . . . . . . . .Th . . . . . . . . . .232.0381* 69. . . . . . . . . Thulium. . . . . . . . . . .Tm . . . . . . . . .168.93421 50. . . . . . . . . Tin . . . . . . . . . . . . . .Sn . . . . . . . . . .118.71 22. . . . . . . . . Titanium . . . . . . . . . .Ti . . . . . . . . . . . 47.88 74. . . . . . . . . Tungsten . . . . . . . . . .W . . . . . . . . . .183.85 112 . . . . . . . Ununbium. . . . . . . . .Uub 116 . . . . . . . Ununhexium . . . . . . .Uuh 110 . . . . . . . Ununnilium . . . . . . . .Uun 114 . . . . . . . Ununquadium. . . . . .Uuq 111 . . . . . . . Unununium. . . . . . . .Uuu. . . . . . . . .272.1535 92. . . . . . . . . Uranium . . . . . . . . . .U. . . . . . . . . . .238.0508* 23. . . . . . . . . Vanadium . . . . . . . . .V . . . . . . . . . . . 50.9415 54. . . . . . . . . Xenon. . . . . . . . . . . .Xe . . . . . . . . . .131.29 70. . . . . . . . . Ytterbium . . . . . . . . .Yb. . . . . . . . . .173.04 39. . . . . . . . . Yttrium . . . . . . . . . . . Y . . . . . . . . . . . 88.90585 30. . . . . . . . . Zinc. . . . . . . . . . . . . .Zn . . . . . . . . . . . 65.39 40. . . . . . . . . Zirconium . . . . . . . . .Zr . . . . . . . . . . . 91.224*Relative atomic mass of the isotope of that element of longest known half-life.Names of individual elements with atomic numbers 110 to 116 are provisional.Based on 2001 IUPAC Table of Standard Atomic Weights of the Elements.Reproduced from Pure Appl. Chem., (2003) Vol. 75, No. 8, pp. 1107-1122 by permission of the copyright owner, IUPAC. ©2003 IUPAC.
Biochemicals
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In 1966, Good, et al. introduced a series of new biological buffers for use in the physiological pH range(1). Prior to that time only a few buffer systems, consisting of weak acid/salt or weak base/salt were available to biologi-cal laboratories. Most of these systems had severe limitations associated with them and are discussed later. The “Good” zwitterionic buffers were designed to meet a genuine need, particularly in the developing field of cellular biology, and have gained rapid acceptance. Zwitterionic buffers, by definition, contain both positive and negative ionizable groups. Secondary and tertiary amines provide the positive charges, while sulfonic and carbox-ylic acid groups provide the negative charges.
There are very few traditional materials which are weak acids or weak bases with pKa values effective between pH 6.0 and 9.0 where most biological processes take place. All of these compounds exhibit some significant devia-tions from the requirements of the “ideal” buffer.
Phosphate is an active participant in many biochemical processes, being an activator, inhibitor and/or a metabolite in many biological systems. Borate buffers complex with a wide variety of important respiratory metabolites and other organic compounds as well. For example, polyols, including carbohy-drates and their derivatives, can significantly lower the pH of borate buffers through complex formation. Phosphates also demonstrate complexing capabilities with divalent and polyvalent cations and can therefore inhibit a series of metal ion-dependent biochemical reactions.
Tris (also a popular traditional biochemical buffer) is a poor buffer below pH 7.5 and is a potentially reactive primary amine which often acts as an inhibitor. It has an appreciable solubility in organic solvents which permits it to penetrate biological membranes and it has a very high pH temperature coefficient.
Bicarbonate is a poor buffer due to the limited solubility of carbon dioxide and the associated problems of liquid/gas equilibrium. Since bicarbonate buffers spontaneously liberate carbon dioxide (equation 1) they require sealed vessels capable of maintaining a carbon dioxide atmosphere in order to sustain a constant pH.
1. CO3= + 2H+ HCO3
- + H+ H2O+CO2
Bicarbonate’s pKa is 6.3 and therefore has a greatly diminished buffering capacity within the physiologically important pH range of 7 to 7.5.
From the Henderson-Hassellbach equation, (equation 2), it can be shown that optimal buffering occurs when the working pH is equal to the pKa of the ionizing group, and falls at an increasing distance from that value. In practice, the useful pH range for a buffer is its pKa ± 1 pH unit.
2. pH = pKa + log [(activity of base) ÷ (activity of acid)]
Good’s buffers were designed to meet the following criteria of an effective “ideal” biological buffer as much as possible:
� pKa between 6.0 and 8.0, the region of most biological reactions� High aqueous solubility� Minimal penetration of biological membranes� Minimal salt effects or effects due to the ionic composition of the solution� Minimal effects due to concentration or temperature� Minimal interactions with mineral cations� Enzymatic and hydrolytic stability� Minimal absorbance between 240 and 700 nm� Easy to purify� Non-toxic� Minimal participation in biochemical reactions
Helpful suggestions1. When preparing concentrated solutions refer to solubility data.2. Media that contain buffers may be adjusted to correct pH with NaOH or
HCl. If this is not feasible, the buffer stock can be adjusted to correct pH before addition to the media.
3. Solutions containing buffers can be sterilized by membrane filtration or normal autoclaving procedures.
4. Don’t autoclave solutions which have been adjusted with Sodium Bicarbonate.
5. PIPES requires the addition of alkali to effect solution.6. Always use pyrogen-free distilled water when preparing media from dry
chemicals.7. Always choose a buffer with a pKa close to the pH that is required. If
the pH is expected to drop during the project, select a buffer with a pKa slightly lower than the pH.
8. Always adjust the pH of the crystalline buffers when placed into solution, otherwise the pH will be incorrect in most instances.
9. Use tetramethylammonium hydroxide when a buffer without mineral cations is required.
10. Whenever possible prepare the buffer solution at the concentration and temperature of the experimental conditions under which it will be used.
11. Other buffers suitable for ultraviolet spectroscopy include many zwitterionic acids. For example, at concentrations of 0.05 M, BES, Bicine, HEPES, MES, PIPES,TES, and Tricine have negligible absorption above 240 nm and can be used successfully at 230 nm. ACES absorbs significantly at 230 nm and ADA is limited to above about 260 nm.
References:1. Good, N. E., Winget, G. D., Winter, W., Connolly, T. N., Izawa, S. and
Singh, R. M. M. (1966) Biochemistry 5, 467-477.2. Schwarzenbach, G. (1955) Helv. Chim. Acta. 38, 1147.3. Good, N. E. and Izawa, S. (1972) Methods Enzymol. 24, 53-68.
Good’s Buffers Guidelines
Biochemicals
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∆ pKa/°C Molarity of Common Formula Temperature Saturated Name Formula Weight Coefficient Solution at 0°C pKa (4°C) pKa (20°C) pKa (37°C)
ACES H2NCOCH2NH 182.20 -0.020 0.22 7.20 6.90 6.54 CH2 CH2SO3H
ADA NH2COCH2N 190.15 -0.011 0.09 6.80 6.60 6.43 (CH2 COOH)2
BES (HOCH2CH2)2NCH2 213.25 -0.016 3.2 7.41 7.15 6.88 CH2 SO3H
Bicine (CH2CH2OH)2NCH2 163.17 -0.018 1.1 8.64 8.35 8.04 COOH
CAPS (C6H11)NH(CH2)3 221.32 -0.021 0.47 10.74 10.40 10.04 SO3H
CHES (C6H11)NH(CH2)2 207.29 -0.011 1.14 9.73 9.55 9.36 SO3H
Glycinamide NH2CH2CONHCH2 132.12 -0.028 1.1 8.85 8.40 7.92 COOH
Glycinamide•HCl H2NCH2CONH2• 110.54 -0.029 4.6 8.66 8.20 7.71 HCl
HEPES HO(CH2)2(C4H8N2) 238.30 -0.014 2.25 7.77 7.55 7.31 (CH2)2SO3H
HEPPS HO3S(CH2)3 252.33 -0.011 1.58 8.18 8.00 7.81 (C4H8N2)(CH2)2OH
MES HO3S(CH2)2 213.25 -0.011 0.65 6.33 6.15 5.96 (C4H8NO)•H2O
MOPS HO3S(CH2)3 209.26 -0.011 3.09 7.38 7.20 7.01 (C4H8NO)
PIPES HO3S(CH2)2(C4H8N2) 302.37 -0.0085 _____ 6.94 6.80 6.66 (CH2)2SO3H
TES (HOCH2)3CNHCH2 229.25 -0.020 2.6 7.82 7.50 7.16 CH2SO3H
TRICINE (CH2OH)3CNHCH2 179.17 -0.021 0.8 8.49 8.15 7.79 COOH
TRIS NH2C(CH2OH)3 121.14 -0.031 2.4 8.80 8.30 7.77
References:1. Good, N. E., Winget, G. D., Winter, W., Connolly, T. N., Izawa, S. and Singh, R. M. M. (1966) Biochemistry 5, 467-477.2. Schwarzenbach, G. (1955) Helv. Chim. Acta. 38, 1147.3. Good, N. E. and Izawa, S. (1972) Methods Enzymol. 24, 53-68.
Physical Properties of Good’s and Other Biological Buffers
Biochemicals
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S-Acetyl Coenzyme A, Lithium SaltULTRAPURE
C23H35N7O17P3SLi3•3H2OFormula Weight: 881.42Product specifications:Form: White powderAssay (Total UV, pH 7.0): ≥ 90%λmax: 259 ± 1 nmεmax (pH 7): [14.6 ± 0.8] x 103
Spectral Ratios: A232/A260: 0.51 ± 0.02 A250/A260: 0.86 ± 0.04 A280/A260: 0.17 ± 0.02Storage: -20°CCAS #75520-41-1
10062 10 mg 25 mg 100 mg 500 mg
N-Acetyl-L-Cysteine
HSCH2CH(NHCOCH3)COOHFormula Weight: 163.19Product specifications:Form: White crystalline powderAssay: 98.0 - 102.0% C5H9NO3S (dry basis)Identity: By IRLoss on Drying: ≤ 1.0%Residue on Ignition: ≤ 0.5%Heavy Metals (Pb): ≤ 10 ppmCAS #616-91-1
10120 100 gm 500 gm 1 kg
Acetylated BSA
See Albumin, Bovine, Acetylated, page 198
AcrylamideULTRAPURE
MB GradeCH2=CHCONH2
Formula Weight: 71.08Product specifications:Form: White/clear crystalline powderAssay: ≥ 99.9%Identity: By IRA (410 nm, 35%, 1 cm): ≤ 0.02A (290 nm, 1%, 1 cm): ≤ 0.1A (280 nm, 7 mM, 1 cm): ≤ 0.04A (252 nm, 7 mM, 1 cm): ≤ 1.20Insoluble Matter (H2O): ≤ 0.005%Insoluble Matter (MeOH): ≤ 0.005%pH (10%, 0.1N NaCl): 5.5 - 7.0Melting Point: 84° ± 1°CAcrylic Acid: ≤ 0.001%Conductivity (35%): ≤ 5 µmhos/cmIron (Fe): ≤ 1 ppmLead (Pb): ≤ 1 ppmGel Performance: Passes testApplications:� Basic component for preparing polyacrylamide
gels� Used at a 19:1 ratio with bis-acrylamideDNA Size Separation by Acrylamide Percentage: DNA Size Range Concentration of (Base Pairs) Acrylamide (%) 100 - 1,000 3.5 80 - 500 5.0 60 - 400 8.0 40 - 200 12.0 10 - 100 20.0CAS #79-06-1
75820 100 gm 500 gm 1 kg
Acrylamide
Reagent GradeC3H5NOFormula Weight: 71.08Product specifications:Form: White, clear crystalline powderAssay: ≥ 99.95%Identity: By IRA (290 nm, 1%, 1 cm): ≤ 0.125pH (10%, H2O): 5.8 - 7.5Acrylic Acid Content: ≤ 0.001%Insoluble Matter: ≤ 0.005%CAS #79-06-1
32800 25 gm 100 gm 500 gm 1 kg
Acrylamide Gel Mixes and Solutions
Glycerol Tolerant Gel (GTG) Buffer, 20X Solution, page 219
RapidGel™ 40% Liquid Acrylamide Stock Solution, page 237
RapidGel-XL 6% DNA Sequencing Gel Solution, page 238
Stop Solution, page 242
Adenine
C5H5N5
Formula Weight: 135.13Product specifications:Form: White to off-white crystalline powderIdentity: By IRAssay: ≥ 98.0%Loss on Drying: ≤ 1.0%Residue on Ignition: ≤ 0.1%Sulfate (SO4): ≤ 0.028%Heavy Metals (Pb): ≤ 20 ppmArsenic (As): ≤ 2 ppmReference constants:λmax (pH 7, NRC): 261 nmεmax (pH 7, NRC): 13.4 x 103
ε260 (pH 7, DBR): 13.3 x 103
CAS #73-24-5
10430 100 gm 1 kg
Adenine Sulfate, Dihydrate
(C5H5N5)2•H2SO4
•2H2OFormula Weight: 404.36Product specifications:Form: White to pale-yellow crystalline powderAssay (Total UV): ≥ 97.0%Identity: By IRSpectral Ratios (pH 1.0): A250/A260: 0.76 ± 0.04 A280/A260: 0.38 ± 0.04Moisture (KF): ≤ 10%CAS #321-30-2
10450 100 gm 1 kg
Biochemicals
189For bulk or alternate pack sizes, email us at [email protected].
Adenosine
C10H13N5O4
Formula Weight: 267.24Product specifications:Form: White crystalline powderAssay (UV): ≥ 98.0%Chromatography (TLC): Homogeneousλmax: 257 nm ± 2 nmSpectral Ratios (0.1N HCl): A250/A260: 0.85 ± 0.03 A280/A260: 0.21 ± 0.02Loss on Drying: ≤ 0.5%Reference constants:λmax (pH 7, NRC): 259 nmεmax (pH 7, NRC): 15.4 x 103
ε260 (pH 6, DBR): 14.9 x 103
CAS #58-61-7
10460 100 gm
Adenosine-5’-Diphosphate, Disodium Salt, Dihydrate(ADP)
C10H13N5O10P2Na2•2H2O
Formula Weight: 507.20Product specifications:Form: White powderAssay (Total UV): 100 ± 4%Spectral Ratios: A250/A260: 0.78 ± 0.03 A280/A260: 0.16 ± 0.02Reference constants:λmax (pH 7, NRC): 259 nmεmax (pH 7, NRC): 15.4 x 103
ε260 (pH 7, DBR): 15.4 x 103
Storage: -20°C in air-tight containers.CAS #16178-48-6
10490 5 gm 10 gm 25 gm
Adenosine-5’-Monophosphate, Free Acid
C10H14N5O7PFormula Weight: 347.22 (anhydrous)* 365.24 (monohydrate)*Product specifications:Form: White crystalline powderAssay (UV, pH 7.0 or 0.01N HCl): 99 ± 3%Identity: By IRSpectral Ratios (pH 7.0): A250/A260: 0.78 ± 0.03 A280/A260: 0.16 ± 0.02 orSpectral Ratios (0.01N HCl): A250/A260: 0.84 ± 0.03 A280/A260: 0.22 ± 0.02Moisture: ≤ 6%* Note: Moisture content may vary from anhydrous to a monohydrate.
Reference constants:λmax (pH 7, NRC): 259 nmεmax (pH 7, NRC): 15.4 x 103
ε260 (pH 7, DBR): 15.3 x 103
CAS #18422-05-4
10520 10 gm 25 gm 100 gm
Adenosine-5’-Monophosphate, Free Acid, Monohydrate(AMP)
C10H14N5O7P•H2OFormula Weight: 365.24Product specifications:Form: White crystalline powderAssay (UV): ≥ 96%λmax: 257 nm ± 2 nmSpectral Ratios (0.1N HCl): A250/A260: 0.84 ± 0.03 A280/A260: 0.22 ± 0.02Loss on Drying: ≤ 6.0%Reference constants:λmax (pH 7, NRC): 259 nmεmax (pH 7, NRC): 15.4 x 103
ε260 (pH 7, DBR): 15.3 x 103
CAS #18422-05-4
10521 25 gm 100 gm 1 kg
Adenosine-5’-Monophosphate, Disodium Salt(AMP)
C10H12N5O7PNa2•nH2O
Formula Weight: 391.19•nH2OProduct specifications:Form: White powderAssay (UV, 0.01N HCl): 98 ± 3%λmax: 257 nm ± 2 nm (0.01N HCl)Spectral Ratios (0.01N HCl): A250/A260: 0.84 ± 0.03 A280/A260: 0.22 ± 0.02Heavy Metals: ≤ 10 ppmReference constants:λmax (pH 7, NRC): 259 nmεmax (pH 7, NRC): 15.4 x 103
ε260 (pH 7, DBR): 15.3 x 103
CAS #4578-31-8
10620 25 gm 100 gm
Adenosine-5’-Triphosphate, 100 mM Solution(ATP)
ULTRAPURE
C10H12N5O13P3Na4
Formula Weight: 595.11Form: 100 mM aqueous solution, pH 7.5Storage: -20°C
77241 25 µmol (250 µl)
Adenosine-5’-Triphosphate, Disodium Salt(ATP)
C10H14N5O13P3Na2•3H2O
Formula Weight: 605.19Product specifications:Form: White crystalline powderAssay (UV, pH 7 or 0.01N HCl): 100 ± 2%λmax: 259 nm ± 2 nm (pH 7.0)εmax (pH 7.0): [15.4 ± 0.3] x 103
Spectral Ratios (pH 7.0): A250/A260: 0.80 ± 0.03 A280/A260: 0.15 ± 0.02 orSpectral Ratios (0.01N HCl): A250/A260: 0.85 ± 0.05 A280/A260: 0.22 ± 0.03Moisture: ≤ 15%Reference constants:λmax (pH 7, NRC): 259 nmεmax (pH 7, NRC): 15.4 x 103
ε260 (pH 7, DBR): 15.4 x 103
Storage: -20°C, protect from moisture.CAS #51963-61-2
10585 5 gm 10 gm 25 gm 100 gm 500 gm
Biochemicals
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S-Adenosyl-L-Methionine Sulfate p-Toluenesulfonate(SAMe-PTS)
C15H23N6O5S(+) n HSO4
(-) n H2SO4C7H7SO3HFormula Weight: 766.80Product specifications:Form: White to off white powderAssay (HPLC): ≥ 95% (dry basis)Moisture (KF): ≤ 2.0%Storage: 4°CCAS #97540-22-2
10601 25 mg 100 mg
Adonitol(D-Ribitol)
C5H12O5
Formula Weight: 152.15Product specifications:Form: White to off-white powderMelting Point: 103° ± 2°CLoss on Drying: ≤ 0.5%Heavy Metals (Pb): ≤ 10 ppmCAS #488-81-3
10635 25 gm 100 gm
ADP
See Adenosine-5’-Diphosphate, Disodium Salt, Dihydrate, page 189
AEBSF Hydrochloride(4-(2-Aminoethyl)-benzenesulfonyl fluoride hydrochloride)
C8H10FNO2S•HClFormula Weight: 239.69Product specifications:Form: Fine white powderAssay (HPLC): ≥ 98%Identity: By IRMelting Point: 181 - 185°CStorage: 4°CCAS #30827-99-7Applications:� Inactivation of trypsin and thrombin(1)
� Synthetic and low molecular weight inhibitor of serum kallikrein(2)
� Specific and irreversible inhibitor of serine prote-ases(3)
Notes: AEBSF irreversibly inactivates thrombin and other serine proteases (e.g. chymotrypsin, kallikrein, plasmin, proteinase K, trypsin) by sulfonylation of a functional group in the active center of the enzyme(1). AEBSF (LD50 determined from the oral dose in mouse is 2.8 gm/kg, which is higher than those for DFP and PMSF) is a substantially less toxic substitute for PMSF and DFP. It is highly water-soluble and has an acidic pH stability. AEBSF is inactivated at 37°C by 50% after 5 hours. We recommend a concentration of stock solution of 20 mM or 100 mM in water or recommended buffer at acidic pH. Stock solutions are stable for up to 2 months at -20°C and for approximately one week at 4°C. AEBSF should be added to an assay buffer just before use at a final concentration of 0.1 - 2 mM(3-5).
References:1. Walsmann, P., Richter, M. and Markwardt, F.
(1972) Acta Biol. Med. Ger. 28, 577-585.2. Markwardt, F., Drawert, J. and Walsmann, P.
(1974) Biochem Pharmacol. 23, 2247-2256.3. Taylor, J. A., Karas, J. L., Ram, M. K., Green, O. M.
and Seidel-Dugan, C. (1995) Mol. Cell. Biol. 8, 4149-4157.
4. Murphy, B. J., Rossie, S., De Jongh, K. S. and Catterall, W. A. (1993) J. Biol. Chem. 268, 27355-27362.
5. Nathan, D. F. and Lindquist, S. (1995) Mol. Cell. Biol. 15, 3917-3925.
11118 200 mg 1 gm
Agar
Product specifications:Form: Light tan granular powderIdentity: Passes testLoss on Drying: ≤ 20.0%Total Ash: ≤ 6.5%CAS #9002-18-0
10654 1 lb 5 lb 25 lb
Agar, BacteriologicalULTRAPURE
Type AProduct specifications:Form: White to cream-colored powderMoisture: ≤ 12%Ash: ≤ 6.5%Gel Strength (1.5%): 550 - 950 gm/cm2
Gel pH (1.5%) after autoclaving: 6.0 - 7.5Gel Point (1.5%): 32 - 38°CMelting Point (1.5%): 80 - 90°CApplication:� Used extensively as a gelling agent in the prepa-
ration of bacteriological culture media. Its main attributes are superior transparency, very reliable reproducibility, and absence of inhibitors capable of hindering optimal development of microorgan-isms. Extraneous matter reduced to a minimum.
CAS #9002-18-0
10906 1 lb 5 lb 25 lb
Agar, Noble AgarULTRAPURE
Product specifications:Form: Pale cream powderMoisture: ≤ 10%Ash: ≤ 1.6%Gel Strength (1.5%): ≥ 700 gm/cm2
Gel pH (1.5%): 5.0 - 7.0Gel Point (1.5%): 32 - 37.5°CMelting Point (1.5%): 80 - 95°CA (430 nm, 1.5%): ≤ 0.150Resistance (1% suspension): ≥ 20,000 OhmsEEO (-mr): ≤ 0.450 (pH 8.4)Application:� Gelling agent essentially free of impurities. Used
mainly in biochemistry and molecular biology procedures wherever increased purity is required.
Typical Agar Preparation: Plates: 15 gm/l of media; 30-35 ml per 90 mm
plate Top Agar: 7 gm/l of media Autoclave for 20 minutes and swirl gently to
evenly distribute the agar. If antibiotics are to be added, allow the agar to cool to 50°C. When pouring plates, gently swirl the medium. A Bunsen burner flame can be passed over the surface of the plates to remove any air bubbles. After the medium has hardened, invert the plates and store at 4°C.
CAS #9002-18-0
10907 100 gm 500 gm 1 kg
Biochemicals
191For bulk or alternate pack sizes, email us at [email protected].
USB Agarose Selection Guide
Genetic Performance Certified (GPC): Functionally tested in specific molecular biology applications (e.g. nucleic acid recovery, in-gel restriction enzyme digestion or ligation) to ensure optimal agarose performance.
Hi-Res: High resolution.
Biochemicals
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Effect of Agarose Concentration on DNA Separation
1X TAE Buffer Gel Concentration 1X TBE Buffer
Size Range (bp) % Size Range (bp)
Agarose - Separation ≥ 500 bp, Genetic Performance Certified [75817]
20,000 - 1,000 0.6 15,000 - 1,000 12,000 - 500 0.8 10,000 - 500 8,000 - 300 1.0 7,000 - 250 6,000 - 200 1.2 5,000 - 200 3,500 - 100 1.5 3,000 - 100 2,000 - 50 2.0 2,000 - 50
Agarose - Hi-Res, Separation ≤ 1000 bp [10132]
1,500 - 100 2.0 1,200 - 100 1,000 - 50 3.0 700 - 40 500 - 20 4.0 200 - 20 300 - 10 5.0 < 100
Agarose - LE [32802]
20,000 - 1,000 0.6 15,000 - 1,000 12,000 - 500 0.8 10,000 - 500 8,000 - 300 1.0 7,000 - 250 6,000 - 200 1.2 5,000 - 200 3,500 - 100 1.5 3,000 - 100 2,000 - 50 2.0 2,000 - 50
Agarose - Low Melt, Separation ≤ 1000 bp, Genetic Performance Certified [32829]
1,500 - 500 2.0 1,000 - 400 700 - 150 3.0 500 - 100 300 - 70 4.0 150 - 10 50 - 10 5.0 < 30
Agarose - Low Melt, Separation ≥ 1000 bp, Genetic Performance Certified [32830]
20,000 - 500 0.75 12,000 - 500 16,000 - 300 1.00 8,000 - 300 10,000 - 250 1.25 4,000 - 200 5,000 - 200 1.50 3,000 - 150 2,500 - 100 1.75 2,000 - 100 1,500 - 50 2.00 1,000 - 50
Agarose - HEULTRAPURE
Product specifications:Form: Granular, free-flowing powderEEO (-mr): 0.23 - 0.26Gel Point (1.5%): 36 ± 1.5°CRemelt Point (1.5%): 88 ± 1.5°CGel Strength (1%): ≥ 750 gm/cm2
Gel Strength (1.5%): ≥ 1200 gm/cm2
Moisture: ≤ 7%Turbidity (1.5%): ≤ 40 NpResidue on Ignition: ≤ 1.0%Sulfate (SO4): ≤ 0.2%Applications:� Counter current electrophoresis� Immunodiffusion
CAS #9012-36-6
32804 100 gm 250 gm 500 gm
Agarose - MEULTRAPURE
Product specifications:Form: Granular, free-flowing powderEEO (-mr): 0.16 - 0.19Gel Point (1.5%): 36 ± 1.5°CRemelt Point (1.5%): 88 ± 1.5°CGel Strength (1%): ≥ 1000 gm/cm2
Gel Strength (1.5%): ≥ 2200 gm/cm2
Moisture: ≤ 7%Turbidity (1.5%): ≤ 40 NpResidue on Ignition: ≤ 0.5%Sulfate (SO4): ≤ 0.2%Applications:� High resolution protein electrophoresis� Immunoelectrophoresis� Electro immunoassays such as rocket assays� Counter current electrophoresisCAS #9012-36-6
32803 25 gm 100 gm 250 gm 500 gm
Biochemicals
193For bulk or alternate pack sizes, email us at [email protected].
ULTRAPURE
MB GradeProduct specifications:Form: Granular, free-flowing powderEEO (-mr): 0.05 - 0.13Gel Point (1.5%): 36.0 ± 1.5°CRemelt Point (1.5%): 88.0 ± 1.5°CGel Strength (1%): ≥ 1,200 gm/cm2
Gel Strength (1.5%): ≥ 2,500 gm/cm2
Moisture: ≤ 7%Turbidity (1.5%): ≤ 40 NpResidue on Ignition: ≤ 0.4%Sulfate (SO4): ≤ 0.14%
Agarose - Separation ≥ 500 bp, Genetic Performance Certified™
* Range of separation depends upon the choice of buffer. Migration in TBE buffer is slower, therefore slightly lower concentrations can be used to obtain the same range of separation.
** Size in base pairs
Guide for separation ranges*Agarose – Separation ≥ 500 bp, Genetic Performance Certified (Product No. 75817) 200 400 600 800 1,000 2,000 4,000 6,000 8,000 10,000 12,000 14,000 16,000 18,000 20,000 bp**
TAE Buffer TBE Buffer
0
Preparation of an Agarose Gel (PNs 75817 & 32802):Weigh out the desired amount of agarose and place in an Erlenmeyer flask with a measured amount of electrophoresis buffer. Example: For an 0.8% gel, add 0.8 gm of agarose and 100 ml of TBE Buffer (1X), to a 200 ml flask. The larger flask ensures against the agarose boiling over. Dissolve the agarose in a boiling water bath or in a revolving-plate microwave oven. All the grains of agarose should be dissolved and the solution clear. Cool the solution to 60°C (70°C for concentrations 2% or above) and pour immediately. Allow the gel to set for one-half hour before using. Make sure to use the same electrophoresis buffer in the gel as for the running buffer.
Related products
TAE Buffer, 10X Solution75904 1 L 5 L
TBE Buffer, 5X Solution75891 1 L 5 L
TBE Buffer, 10X Ready-Mixed Powder70454 6 x 200 ml
Genetic performance tests: DNase: Passes test RNase: Passes test Restriction Enzyme Digestion: Passes test Ligation: Passes test DNA Binding: Passes test Blotting: Passes test Gel Background: Passes test DNA Resolution: Passes testSeparation guidelines:Range of separation in 1X TAE: 0.6%: 20,000-1,000 bp 0.8%: 12,000-500 bp 1.0%: 8,000-300 bp 1.2%: 6,000-200 bp 1.5%: 3,500-100 bp 2.0%: 2,000-50 bp
Range of separation in 1X TBE: 0.6%: 15,000-1,000 bp 0.8%: 10,000-500 bp 1.0%: 7,000-250 bp 1.2%: 5,000-200 bp 1.5%: 3,000-100 bp 2.0%: 2,000-50 bpApplications:� Separation of DNA or RNA fragments ≥ 500 bp.� PCR analysis.� Recovery of separated DNA or RNA fragments
from gel.� Blotting applications such as Southerns and
Northerns.CAS #9012-36-6
75817 25 gm 100 gm 250 gm 500 gm
Biochemicals
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* Range of separation depends upon the choice of buffer. Migration in TBE buffer is slower, therefore slightly lower concentrations can be used to obtain the same range of separation.
** Size in base pairs
Guide for separation ranges*Agarose – LE (Product No. 32802)
200 400 600 800 1,000 2,000 4,000 6,000 8,000 10,000 12,000 14,000 16,000 18,000 20,000 bp**
TAE Buffer TBE Buffer
0
ULTRAPURE
MB GradeProduct specifications:Form: Granular, free-flowing powderEEO (-mr): 0.05 - 0.13Gel Point (1.5%): 36 ± 1.5°CRemelt Point (1.5%): 88 ± 1.5°CGel Strength (1%): ≥ 1,200 gm/cm2
Gel Strength (1.5%): ≥ 2,500 gm/cm2
Moisture: ≤ 7%Turbidity (1.5%): ≤ 40 NpResidue on Ignition: ≤ 0.4%Sulfate (SO4): ≤ 0.14%DNase: Passes testRNase: Passes testGel Background: Passes test
Agarose - LE
Separation guidelines:Range of separation in 1X TAE: 0.6%: 20,000-1,000 bp 0.8%: 12,000-500 bp 1.0%: 8,000-300 bp 1.2%: 6,000-200 bp 1.5%: 3,500-100 bp 2.0%: 2,000-50 bpRange of separation in 1X TBE: 0.6%: 15,000-1,000 bp 0.8%: 10,000-500 bp 1.0%: 7,000-250 bp 1.2%: 5,000-200 bp 1.5%: 3,000-100 bp 2.0%: 2,000-50 bp
Applications:� Routine separation of DNA or RNA fragments.� PCR analysis.� Blotting applications such as Southerns and
Northerns.� Protein analysis such as immunodiffusion, rocket
assays, and immuno electrophoresis.CAS #9012-36-6
32802 25 gm 100 gm 250 gm 500 gm 1 kg
Biochemicals
195For bulk or alternate pack sizes, email us at [email protected].
* Range of separation depends upon the choice of buffer. Migration in TBE buffer is slower, therefore slightly lower concentrations can be used to obtain the same range of separation.
** Size in base pairs
Guide for separation ranges*Agarose – Hi-Res, Separation ≤ 1,000 bp (Product No. 10132) 10 20 30 40 50 60 70 80 90 100 200 300 400 500 600 700 800 900 1,000 1,100 1,200 1,300 1,400 1,500 bp**
TAE Buffer TBE Buffer
0
ULTRAPURE
MB GradeProduct specifications:Form: Granular, free-flowing powderEEO (-mr): ≤ 0.12Gel Point (3%): ≤ 35.5°CRemelt Point (3%): ≤ 80°CGel Strength (1.5%): ≥ 600 gm/cm2
Turbidity (1.5%): ≤ 50 NpGel Strength (3%): ≥ 1,500 gm/cm2
Moisture: ≤ 7%Residue on Ignition: ≤ 0.35%
Agarose - Hi-Res, Separation ≤ 1000 bp
Sulfate (SO4): ≤ 0.11%DNase: Passes testRNase: Passes testSeparation guidelines:Range of separation in 1X TAE: 1.8%: 1,000-500 bp 2.0%: 1,500-100 bp 3.0%: 600-150 bp 4.0%: 500-20 bp 4.5%: 350-50 bp 5.0%: 300-10 bp
Range of separation in 1X TBE: 2.0%: 1,200-100 bp 3.0%: 700-40 bp 4.0%: 200-20 bp 5.0%: < 100 bpApplications:� Highly resolved separation of DNA or RNA frag-
ments ≤ 1000 bp� PCR analysisCAS #9012-36-6
10132 25 gm 100 gm 500 gm
Related products – Buffers & stock biochemicals:
Ethidium Bromide32813 1 gm 5 gm 25 gm
Ethidium Bromide Drops75816 5 ml
TAE Buffer, 10X Solution75904 1 L 5 L
TBE Buffer, 5X Solution75891 1 L 5 L
TBE Buffer, 10X Ready-Mixed Powder70454 6 x 200 ml
Related products – DNA markers:
PCR Markers, 50 – 2000 bp76710 250 µl
DNA Ladder, 100 bp76712 500 µl
DNA Ladder, 1 kb Plus76714 500 µl
Biochemicals
196 888-362-2447 | 216-765-5000 | usb.affymetrix.com
ULTRAPURE
MB GradeProduct specifications:Form: Granular, free-flowing powderEEO (-mr): ≤ 0.12Gel Point (1.5%): 24 - 28°CRemelt Point (1.5%): ≤ 65.5°CGel Strength (1%): ≥ 250 gm/cm2
Gel Strength (1.5%): ≥ 500 gm/cm2
Moisture: ≤ 7%Turbidity (1.5%): ≤ 40 NpResidue on Ignition: ≤ 0.4%Sulfate (SO4): ≤ 0.12%
Agarose - Low Melt Separation ≥ 1000 bp, Genetic Performance Certified
Genetic performance tests: DNase: Passes test RNase: Passes test In-Gel Restriction Enzyme Digestion: Passes test In-Gel Ligation: Passes test DNA Binding: Passes test Gel Background: Passes test DNA Resolution: Passes test Agarase Digestion: Passes testSeparation guidelines:Range of separation in 1X TAE: 0.75%: 20,000 - 500 bp 1.00%: 16,000 - 300 bp 1.25%: 10,000 - 250 bp 1.50%: 5,000 - 200 bp 1.75%: 2,500 - 100 bp 2.00%: 1,500 - 50 bp
Range of separation in 1X TBE: 0.75%: 12,000 - 500 bp 1.00%: 8,000 - 300 bp 1.25%: 4,000 - 200 bp 1.50%: 3,000 - 150 bp 1.75%: 2,000 - 100 bp 2.00%: 1,000 - 50 bpApplications:� Enzymatic modification of nucleic acids in re-
melted gels.� Separation of DNA or RNA fragments ≥ 1,000 bp.� In-gel recovery of nucleic acid fragments.� PCR analysis.� Labile sample procedures.� Suitable for cell culture and cloning applications.CAS #9012-36-6
32830 25 gm 100 gm
* Range of separation depends upon the choice of buffer. Migration in TBE buffer is slower, therefore slightly lower concentrations can be used to obtain the same range of separation.
** Size in base pairs
Guide for separation ranges*Agarose – Low Melt Separation ≥ 1,000 bp, Genetic Performance Certified (Product No. 32830)
Isolating DNA from low-melting temperature agarose (PNs 32829 & 32830):Follow procedure for preparing an agarose gel and be sure to run the electrophoresis at 4°C to keep the gel from melting. Excise the DNA band of interest and transfer it to a microcentrifuge tube. Cut as close to the DNA band as possible to avoid any unnecessary agarose. Add 5 volumes of 20 mM Tris-HCl (pH 8.0), 1 mM EDTA (pH 8.0) to the tube with the agarose. Cap the tube and heat for 5 minutes at 65°C. The aga-rose should melt. Cool the solution to room temperature and add an equal volume of equilibrated phenol. Vortex for 20 seconds and spin at 400 x g for 10 minutes (20°C). Remove the aqueous phase (top) to a new microcentrifuge tube. Re-extract with phenol:chloroform and then with chloroform. Precipitate the final aqueous phase with 0.2 volumes of 10 mM ammonium acetate and 2 volumes of ethanol. Recover the DNA by centrifugation.
100 200 300 400 500 600 700 800 900 1,000 2,000 4,000 6,000 8,000 10,000 12,000 14,000 16,000 18,000 20,000 bp**0
TAE Buffer TBE Buffer
Related products
TAE Buffer, 10X Solution75904 1 L 5 L
TBE Buffer, 5X Solution75891 1 L 5 L
TBE Buffer, 10X Ready-Mixed Powder70454 6 x 200 ml
Biochemicals
197For bulk or alternate pack sizes, email us at [email protected].
ULTRAPURE
MB GradeProduct specifications:Form: Granular, free-flowing powderEEO (-mr): ≤ 0.12Gel Point (4%): ≤ 35°CRemelt Point (4%): ≤ 65°CGel Strength (4%): ≥ 1,000 gm/cm2
Moisture: ≤ 5%Residue on Ignition: ≤ 0.3%Sulfate (SO4): ≤ 0.12%
Agarose - Low Melt Separation ≤ 1000 bp, Genetic Performance Certified
Genetic performance tests: DNase: Passes test RNase: Passes test In-Gel Restriction Enzyme Digestion: Passes test In-Gel Ligation: Passes test DNA Binding: Passes test Gel Background: Passes test DNA Resolution: Passes test Agarase Digestion: Passes testSeparation guidelines:Range of separation in 1X TAE: 2.0%: 1,500-100 bp 3.0%: 700-150 bp 4.0%: 300-70 bp 5.0%: 50-10 bp
Range of separation in 1X TBE: 2.0%: 1,000-400 bp 3.0%: 500-100 bp 4.0%: 150-10 bp 5.0%: < 30 bpApplications:� Enzymatic modification of nucleic acids in re-
melted gels.� Separation of DNA or RNA fragments ≤ 1,000 bp.� In-gel recovery of nucleic acid fragments.� PCR analysis.� Labile sample procedures.CAS #9012-36-6
32829 25 gm 100 gm
* Range of separation depends upon the choice of buffer. Migration in TBE buffer is slower, therefore slightly lower concentrations can be used to obtain the same range of separation.
** Size in base pairs
Guide for separation ranges*Agarose – Low Melt Separation ≤ 1,000 bp, Genetic Performance Certified (Product No. 32829) 10 20 30 40 50 60 70 80 90 100 200 300 400 500 600 700 800 900 1,000 1,100 1,200 1,300 1,400 1,500 bp**
TAE Buffer TBE Buffer
0
ULTRAPURE
MB GradeProduct specifications:Form: Granular, free-flowing powderEEO (-mr): ≤ 0.12Gel Point (1.5%): ≤ 20°CRemelt Point (1.5%): ≤ 63°C
Agarose - Ultra Low Melt
Gel Strength (1.5%): ≥ 300 gm/cm2
Moisture: ≤ 7%Turbidity (1.5%): ≤ 40 NpResidue on Ignition: ≤ 0.35%Sulfate (SO4): ≤ 0.14%DNase: Passes testRNase: Passes test
Applications:� Labile sample procedures.� Cell culture and cloning applications (hybrid-
omas).CAS #9012-36-6
32821 10 gm 25 gm
Biochemicals
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β-Alanine
NH2CH2CH2COOHFormula Weight: 89.09Product specifications:Form: White crystalline powderAssay: ≥ 98.0% (as is)Identity: By IRChromatography (TLC): HomogeneousLoss on Drying: ≤ 0.2%Residue on Ignition: ≤ 0.2%Ammonia (NH4): ≤ 0.1%Chloride (Cl): ≤ 0.04%Sulfate (SO4): ≤ 0.05%CAS #107-95-9
10665 200 gm 1 kg 5 kg
Albumin, Bovine, Protease-Free
MB GradeProtease FreeSource: Bovine BloodProduct specifications:Form: Lyophilized powderAssay (electrophoresis): ≥ 98.0%Protein: ≥ 96.0%pH (1%, 0.15 M NaCl): 6.8 - 7.2Moisture: ≤ 5.0%Residue on Ignition: ≤ 1.5%IgG: Passes testHeavy Metals (Pb): ≤ 10 ppmProtease: Passes testStorage: 4°CCAS #9048-46-8
10867 10 gm 50 gm 100 gm
Tired of columns? Try ExoSAP-IT reagent, our enzymatic PCR cleanup solution.
Albumin, Bovine, Acetylated(Acetylated BSA)
ULTRAPURE
MB GradeSource: Bovine BloodProduct specifications:Form: Clear, colorless solution in deionized water;
packed under aseptic, nuclease-free conditions.Assay: Single band by SDS-polyacrylamide gel elec-
trophoresisProtein Concentration: 25 - 50 mg/mlContaminant Testing: DNase (endo): Passes test DNase (exo): Passes test RNase: Passes test Protease: Passes testFunctional Test: No inhibition by albumin on diges-
tion of lambda DNA.Form: Many nucleases are irreversibly inactivated by
acetylation, but the stabilizing action of BSA is unaffected by this procedure.(1,2) Thus, acetylation ensures that the use of albumin does not intro-duce additional nucleases to reaction mixture.
Solubility: Soluble in waterApplication:� Bovine Serum Albumin is used extensively by
molecular biologists to stabilize labile enzymes such as restriction endonucleases, polymerases, and ligases.
References:1. Epstein, C. J. and Goldberger, R. F. (1964) J. Biol.
Chem. 239, 1087-1089.2. Lackey, D. and Linn, S. (1980) Meth. in Enzymol.,
65, 26-28.Storage: -20°CCAS #9048-46-8
10848 25 mg 125 mg
Albumin, Bovine, Crystallized, pH 5.2
Source: Bovine BloodProduct specifications:Form: Crystallized powderAssay (electrophoresis): ≥ 99%Identity: By IRProtein: ≥ 98%pH (7%, 0.15 M NaCl): 5.0 - 5.4Moisture (KF) when packaged: ≤ 5.0%Sodium (Na): 0 - 5.0 mg/gmChloride (Cl): 0 - 3.0 mg/gmStorage: -20°CCAS #9048-46-8
10856 5 gm 10 gm 100 gm
Albumin, Bovine, Cohn Fraction V, pH 5.2(Cold Alcohol Fractionated)
Source: Bovine BloodProduct specifications:Form: Lyophilized powderAssay (electrophoresis): ≥ 96%Protein: ≥ 95%pH (1%, H2O): 5.0 - 5.4Residue on Ignition: ≤ 1.5%Loss on Drying: ≤ 6.0%Heavy Metals (Pb): ≤ 50 ppmStorage: 4°C, desiccatedCAS #9048-46-8
10852 50 gm 100 gm 500 gm
Albumin, Bovine, pH 7(Heat-Shock Fractionated)
Source: Bovine BloodProduct specifications:Form: Lyophilized powderAssay (electrophoresis): ≥ 98%Protein: ≥ 96.0%pH (2%, H2O): 6.5 - 7.5Loss on Drying: ≤ 5.0%Residue on Ignition: ≤ 2.0%Storage: 4°CCAS #9048-46-8
10857 50 gm 100 gm 500 gm 1 kg
Albumin, Bovine, RIA Grade, pH 7
Source: Bovine BloodProduct specifications:Form: Lyophilized powderAssay (electrophoresis): ≥ 98.0%Protein: ≥ 96.0%pH (1%. 0.15 M NaCl): 6.8 - 7.2Loss on Drying: ≤ 5.0%Residue on Ignition: ≤ 2.0%IgG: NegativeInsulin: NegligibleFolic Acid: NegligibleT3, T4: NegligibleApplication:� Suitable for use in radioimmuno assayStorage: 4°CCAS #9048-46-8
10868 10 gm 50 gm 100 gm
Biochemicals
199For bulk or alternate pack sizes, email us at [email protected].
Albumin, Egg(Ovalbumin)
Source: Egg White, Spray DriedProduct specifications:Form: Off-white crystalsProtein (as is): ≥ 80%pH: 6.5 - 8.0Moisture: ≤ 8.0%Salmonella: NegativeCAS #9006-59-1
15460 1 lb 5 lb 25 lb
Albumin, Egg(Ovalbumin)
Source: Chicken EggProduct specifications:Form: White to pale yellow freeze dried powderAssay: ≥ 98.0% (as confirmed by SDS-PAGE)Moisture: ≤ 6.0%Solubility: Soluble in distilled water or dilute bufferStorage: -20°CCAS #9006-59-1
10865 5 gm
Alcohol Dehydrogenase
(E.C.1.1.1.1)Source: YeastProduct specifications:Form: Lyophilized powderActivity: ≥ 300 units/mg proteinUnit Definition: One unit reduces 1 µmol NAD/min-
ute at 25°C, pH 8.8.Contaminating Activities:Carboxylase: ≤ 0.05%GAPDH: ≤ 0.05%PGK: ≤ 0.05%MK: ≤ 0.03%Aldolase: ≤ 0.02%PK: ≤ 0.01%LDH: ≤ 0.01%Storage: -20°CCAS #9031-72-5
10895 75,000 units
Aminoacetic Acid
See Glycine, page 219
4-Amino Antipyrene
C11H13N3OFormula Weight: 203.24Product specifications:Form: Pale yellow to light amber crystalline powderAssay: ≥ 98.0%Identity: By IRSolubility (5%, H2O): Clear solutionMelting Point: 109 ± 2°CCAS #83-07-8
11056 50 gm 100 gm
p-Amino Benzoic Acid (PABA)
USBioAnalyzedC7H7NO2
Formula Weight: 137.14Product specifications:Form: White to off-white crystalline powderAssay (dry basis): 98.5 - 101.5%Identity (A, B): Passes testsMelting Point: 186 - 189°CLoss on Drying: ≤ 0.2%Residue on Ignition: ≤ 0.1%Heavy Metals (Pb): ≤ 0.002%Volatile Diazotizable Substances: ≤ 0.002%Ordinary Impurities: Passes testCAS #150-13-0
11055 200 gm 500 gm 1 kg 5 kg
L-α-Amino-N-Butyric Acid
CH3CH2CH(NH2)COOHFormula Weight: 103.12Product specifications:Form: White crystalline powderAssay: ≥ 98.0% (as is)Identity: By IRChromatography (TLC): HomogeneousMoisture (KF): ≤ 1%CAS #1492-24-6
11079 1 gm 5 gm
2-Aminoethane Sulfonic Acid
See Taurine, page 243
L-α-Aminoglutaric Acid
See L-Glutamic Acid, page 218
6-Amino Purine
See Adenine, page 188
Ammonium AcetateULTRAPURE
ACS Reagent GradeCH3COONH4
Formula Weight: 77.08Product specifications:Form: White/clear crystalsAssay: ≥ 97.0%pH (5%, H2O): 6.7 - 7.3Insoluble Matter: ≤ 0.005%Residue on Ignition: ≤ 0.01%Chloride (Cl): ≤ 5 ppmNitrate (NO3): ≤ 0.001%Sulfate (SO4): ≤ 0.001%Heavy Metals (Pb): ≤ 5 ppmIron (Fe): ≤ 5 ppmSolubility (5 M, H2O): Clear, colorlessApplications:� Used in the precipitation of DNA� Said to help decrease the co-precipitation of
dNTPs� Asymmetric PCR precipitationStock Solution, 7.5 M: Ammonium Acetate: 289 gm dH2O:175 ml Adjust volume with water to 500 ml. Filter sterilize.CAS #631-61-8
11251 500 gm 1 kg
Ammonium Acetate (NH4OAc), 5 M SolutionULTRAPURE
MB GradeForm: 5 M solution of ammonium acetate (NH4OAc)
in high purity dH2O. Solution is 0.2 µm filtered and dispensed into durable, square Nalgene bottles.
Contaminant Testing: DNase (endo): Passes test DNase (exo): Passes test RNase: Passes test
75901 100 ml 500 ml
Biochemicals
200 888-362-2447 | 216-765-5000 | usb.affymetrix.com
Ammonium Persulfate (APS)(Ammonium Peroxydisulfate)
ULTRAPURE
ACS Reagent Grade(NH4)2S2O8
Formula Weight: 228.19Product specifications:Form: Off-white crystalline powderAssay: ≥ 98.0%Residue on Ignition: ≤ 0.05%Insoluble Matter: ≤ 0.005%Titratable Free Acid: ≤ 0.04 meq/gmChloride and Chlorate (Cl): ≤ 0.001%Heavy Metals (Pb): ≤ 0.005%Iron (Fe): ≤ 0.001%Manganese (Mn): ≤ 0.5 ppmApplication:� A high speed initiator used to catalyze acrylamide
polymerizationWorking Stock Solution (10%): Ammonium Persulfate: 1 gm dH2O: to 10 ml This solution should be stored at 4°C and pre-
pared fresh weekly.CAS #7727-54-0
76322 10 gm 100 gm 1 kg
Ammonium SulfateULTRAPURE
ACS Reagent Grade(NH4)2SO4
Formula Weight: 132.14Product specifications:Form: White crystalline powderAssay: ≥ 99.0%pH (5%, H2O, 25°C): 5.0 - 6.0Insoluble Matter: ≤ 0.005%Residue on Ignition: ≤ 0.005%Chloride (Cl): ≤ 5 ppmNitrate (NO3): ≤ 0.001%Phosphate (PO4): ≤ 5 ppmHeavy Metals (Pb): ≤ 5 ppmIron (Fe): ≤ 5 ppmApplication:� Used in the purification of enzymes and other
proteins by precipitationCAS #7783-20-2
11254 1 kg 5 kg
AMP
See Adenosine-5’-Monophosphate, Disodium Salt, page 189
Ampicillin, Sodium SaltULTRAPURE
USBioAnalyzedC16H18N3NaO4SFormula Weight: 371.39Product specifications:Form: White to off-white powderActivity: 845 µg - 988 µg of C16H19N3O4S/mg (dry
basis)Activity: 828 µg - 988 µg of C16H19N3O4S/mg (as is)Identity (A, B): Passes testspH (10 mg/ml): 8.0 - 10.0Moisture (KF): ≤ 2.0%Dimethylaniline: Meets requirementsMethylene chloride: ≤ 0.2%Storage: Controlled room temperature, in tightly
closed containers.CAS #69-52-3
11259 5 gm 25 gm 100 gm
Antibiotic G-418 Sulfate
See G-418 Sulfate, page 217
AprotininULTRAPURE
Source: Bovine LungC284H432N84O79S7
Formula Weight: 6511.44Product specifications:Form: Lyophilized powderActivity: 5,100 - 7,300 Kallikrein Inhibitor units/mg
proteinPurity (gel filtration): ≥ 95%Loss on Drying: ≤ 6.0%Applications:� A useful protease inhibitor against kallikrein,
trypsin, plasmin and chymotrypsin� Not effective against papainWorking Concentration: 1-2 mg/ml Soluble in 0.01 M HEPES (pH 8.0)Note: Avoid repeated freezing and thawing of stock
solutions.Storage: 4°C in air-tight containers.CAS #9087-70-1
11388 10 mg 25 mg 100 mg
L- (+)-Arabinose
C5H10O5
Formula Weight: 150.13Product specifications:Form: White to off-white crystalline powderAssay (GC): ≥ 96%Loss on Drying: ≤ 0.5%Residue on Ignition: ≤ 0.5%Chloride (Cl): ≤ 100 ppmSulfate (SO4): ≤ 100 ppmArsenic (As): ≤ 3 ppmIron (Fe): ≤ 50 ppmHeavy Metals (Cu): ≤ 5 ppmCAS #5328-37-0
11405 25 gm 100 gm 1 kg
L-Arabinose
High Purity GradeC5H10O5
Formula Weight: 150.13Product specifications:Form: White crystalline powder[α]20
D : +103.0° ± 1.5° (c=4 in H2O, NH3)Loss on Drying: ≤ 0.5%Residue on Ignition: ≤ 0.1%Arsenic (As): ≤ 0.5 ppmIron (Fe): ≤ 5 ppmHeavy Metals (Pb): ≤ 10 ppmCAS #5328-37-0
11406 25 gm 100 gm 1 kg
L-Arginine
C6H14N4O2
Formula Weight: 174.20Product specifications:Form: White crystalline powderAssay: ≥ 98.5%Identity: By IR[α]20
D : +26.9° to +27.9° (c=8 in 6N HCl)Loss on Drying: ≤ 0.5%Residue on Ignition: ≤ 0.3%Heavy Metals (Pb): ≤ 15 ppmAmmonium (NH4): ≤ 0.02%Arsenic (As2O3): ≤ 1 ppmChloride (Cl): ≤ 0.05%Iron (Fe): ≤ 30 ppmSulfate (SO4): ≤ 0.03%CAS #74-79-3
11490 100 gm 500 gm 1 kg
Biochemicals
201For bulk or alternate pack sizes, email us at [email protected].
L-Arginine, Hydrochloride
NH2C(NH)NH(CH2)3CH(NH2)COOH•HClFormula Weight: 210.66Product specifications:Form: White crystalline powderAssay: 98.5 - 101.5%Chromatography (TLC): HomogeneousIdentity: By IR[α]20
D : +21.4° to +23.6° (c=8 in 6N HCl)Loss on Drying: ≤ 0.20%Residue on Ignition: ≤ 0.10%Chloride (Cl): 16.5 to 17.1%Sulfate (SO4): ≤ 0.03%Heavy Metals (Pb): ≤ 20 ppmOrganic Volatile Impurities: Meets requirementsCAS #1119-34-2
11500 100 gm 500 gm 1 kg
L-Ascorbic Acid(Vitamin C)
USBioAnalyzedC6H8O6
Formula Weight: 176.12Product specifications:Form: White to slightly yellow crystalline powderAssay: 99.0 - 100.5%Identity A: By KBrIdentity B: Passes test[α]25
D : +20.5° to +21.5° (c=10 in H2O)Residue on Ignition: ≤ 0.1%Heavy Metals (Pb): ≤ 0.002%CAS #50-81-7
11545 1 kg 5 kg
L-Asparagine, Anhydrous
NH2COCH2CH(NH2)COOHFormula Weight: 132.12Product specifications:Form: White crystalline powderAssay: ≥ 98.5%Chromatography (TLC): HomogeneousIdentity: By IR[α]20
D : +33° to +36° (c=2 in 10% HCl)Loss on Drying: ≤ 1.0%CAS #70-47-3
11590 100 gm 500 gm 1 kg
L-Asparagine, Monohydrate
C4H8N2O3•H2O
Formula Weight: 150.13Product specifications:Form: White crystalline powderAssay: ≥ 98.0% (as is)Identity: By IRChromatography (TLC): Homogeneous[α]20
D : +33° to +36° (c=10 in 3N HCl)Loss on Drying: ≤ 12.5%Residue on Ignition: ≤ 0.1%Heavy Metals (Pb): ≤ 10 ppmArsenic (As): ≤ 3 ppmIron (Fe): ≤ 20 ppmSulfate (SO4): ≤ 0.025%Lead (Pb): ≤ 5 ppmCAS #5794-13-8
11595 100 gm 1 kg 5 kg
L-Aspartic Acid
C4H7NO4
Formula Weight: 133.10Product specifications:Form: White crystalline powderAssay: ≥ 98.0%Identity: By IRChromatography (TLC): Homogeneous[α]20
D : +24.5° to +26.0° (c=8 in 6N HCl)Loss on Drying: ≤ 0.25%Residue on Ignition: ≤ 0.1%Heavy Metals (Pb): ≤ 10 ppmArsenic (As): ≤ 3 ppmCAS #56-84-8
11620 100 gm 1 kg 5 kg
L-Aspartic Acid, Sodium Salt, Monohydrate(Sodium Aspartate)
NH2CH(COOH)CH2COONa•H2OFormula Weight: 173.10Product specifications:Form: White crystalline powderAssay: ≥ 98.0% (dry basis)Identity: By IRChromatography (TLC): Homogeneous[α]20
D : +18° to +21° (c=8 in 6N HCl, dry basis)Loss on Drying: ≤ 0.5%Heavy Metals (Pb): ≤ 10 ppmArsenic (As): ≤ 1 ppmChloride (Cl): ≤ 0.05%Iron (Fe): ≤ 30 ppmSulfate (SO4): ≤ 0.05%CAS #323194-76-9
11635 1 kg
L-Aspartic Acid, Magnesium Salt, Dihydrate
(C4H6NO4)2Mg•2H2OFormula Weight: 324.53 (dihydrate)Product specifications:Form: White powderAssay: ≥ 95.0% (dry basis)[α]20
D : +18.0° to +23.0° (20°C, c=2 in 18% HCl)Moisture (KF): ≤ 17%Heavy Metals (Pb): ≤ 10 ppmArsenic (As2O3): ≤ 1 ppmChloride (Cl): ≤ 0.021%Sulfate (SO4): ≤ 0.050%Iron (Fe): ≤ 30 ppmCAS #2068-80-6
11631 100 gm 1 kg
ATP
See Adenosine-5’-Triphosphate, page 189
Avidin-Alkaline Phosphatase Conjugate
See Streptavidin-Alkaline Phosphatase Conjugate, page 242
Biochemicals
202 888-362-2447 | 216-765-5000 | usb.affymetrix.com
Bacitracin
USBioAnalyzedC66H103N17O16SFormula Weight: 1422.69Product specifications:Form: White to pale buff powderActivity: ≥ 40 units of bacitracin/mg (as is)Identity: Passes testpH (10,000 units/ml): 5.5 - 7.5Loss on Drying: ≤ 5.0%Residue on Ignition: ≤ 0.5%CAS #1405-87-4
11805 5 gm 25 gm
Bacterial Culture Media
LB Agar, Ready-Made Powder, page 224LB Broth, Ready-Made Powder, page 224Luria Agar, Ready-Made Powder, page 225Luria Broth, Ready-Made Powder, page 225NZCYM Broth, Ready-Made Powder, page 231Terrific Broth, Ready-Made Powder, page 2452X YT Broth, page 251
Bacteriological Peptone(Peptones)
ULTRAPURE
Product specifications:Form: PowderMoisture: ≤ 5.0%Ash: 4.0 - 6.65%pH (6% solution): 6.6 - 7.8Amino Nitrogen: ≥ 5.1%Total Nitrogen: ≥ 12%Protein (N x 6.38): ≥ 76%An extremely high grade of peptone prepared spe-
cifically for general bacteriological purposes.CAS #73049-73-7
20048 1 lb 5 lb 25 lb
BCIP
See 5-Bromo-4-Chloro-3-Indolyl Phosphate, page 203
Betaine, 5 M SolutionULTRAPURE
MB GradeForm: 5 M solution of betaine in deionized dH2O.Contaminant Testing: DNase (endo): Passes test DNase (exo): Passes test RNase: Passes testFunctional Test: Functionally tested for use in PCR.Storage: -20°C
77507 1.5 ml 5 x 1.5 ml 10 ml
Bicine(N,N-bis[2-Hydroxyethyl]glycine)
(CH2CH2OH)2NCH2COOHFormula Weight: 163.17Product specifications:Form: White/clear crystalline powderAssay: 98.5 - 101.5% (dry basis)Identity: By IRpKa (20°C): 8.35 ± 0.15A (250 nm, 10%, 1 cm): ≤ 0.1Loss on Drying: ≤ 0.5%Iron (Fe): ≤ 30 ppmLead (Pb): ≤ 10 ppmCAS #150-25-4
12091 100 gm 1 kg
Bilirubin
C33H36N4O6
Formula Weight: 584.66Product specifications:Form: Bright orange fine powderAssay: ≥ 97.5% (By UV, Emax = 61.2 x 103, as is)Identity: By IRMoisture (KF): ≤ 2.0%λmax: 453 ± 2 nmStorage: Store protected from light.CAS #635-65-4
12110 1 gm 5 gm
Bis-Acrylamide
See N, N’-Methylene-bis-Acrylamide, page 228
Bis-Tris(bis[2-Hydroxyethyl]imino-tris[hydroxymethyl]methane)
ULTRAPURE
(HOCH2CH2)2NC(CH2OH)3
Formula Weight: 209.24Product specifications:Form: White/clear crystalline powderAssay: ≥ 98.5% (dry basis)Identity: By IRpKa (20°C): 6.50 ± 0.15A (280 nm, 1 M, 1 cm): ≤ 0.4Loss on Drying: ≤ 0.5%Iron (Fe): ≤ 30 ppmLead (Pb): ≤ 10 ppmCAS #6976-37-0
12112 100 gm 500 gm 1 kg
Blotting Reagents
Denhardt’s Solution, 50X, page 209SSC, 20X Solution, page 241SSPE, 20X Solution, page 241TBS, 20X Solution, pH 7.4, page 244TBS with Tween (TBST), 20X Solution, pH 7.4,
page 244
Boric AcidULTRAPURE
MB GradeH3BO3
Formula Weight: 61.83Product specifications:Form: White crystalline powderAssay: ≥ 99.5%Insoluble in MeOH: ≤ 0.005%Nonvolatile with MeOH: ≤ 0.05%Sulfate (SO4): ≤ 0.01%Heavy Metals (Pb): ≤ 0.001%Arsenic (As): ≤ 1 ppmCalcium (Ca): ≤ 0.005%Chloride (Cl): ≤ 0.001%Iron (Fe): ≤ 0.001%Phosphate (PO4): ≤ 0.001%Functional Test: Meets or exceeds criteria for high
quality sequencing gel electrophoresis with Sequenase T7 DNA Polymerase.
DNase (endo): Passes testDNase (exo): Passes testRNase: Passes testProtease: Passes testCAS #10043-35-3
76324 500 gm 1 kg
Biochemicals
203For bulk or alternate pack sizes, email us at [email protected].
Brewers Yeast
See Yeast, Brewers, page 251
Brilliant Blue G(Coomassie® Brilliant Blue G-250)
ULTRAPURE
(C.I.42655)C47H48N3S2O7NaFormula Weight: 854.02Product specifications:Form: Dark blue/purple crystalline powderDye content: Approx. 60%Loss on Drying: ≤ 8%λmax: 607 nm ± 1 nmεmax: 38 x 103
Application:� Used for quantitative staining of proteins on poly-
acrylamide electrophoresis gels.CAS #6104-58-1
32812 25 gm 100 gm
Brilliant Blue R(Coomassie Brilliant Blue R-250)
ULTRAPURE
(C.I.42660)C45H44N3O7S2NaFormula Weight: 825.97Product specifications:Form: Dark purple/violet powderIdentity: By IRλmax: 586 ± 2 nmεmax: ≥ 45.0 x 103
Moisture (KF): ≤ 6.0%Application:� Used for quantitative staining of proteins on poly-
acrylamide electrophoresis gels.CAS #6104-59-2
32826 25 gm 100 gm
5-Bromo-4-Chloro-3-Indolyl-α-D-Galactopyranoside(X-α-GAL)
ULTRAPURE
C14H15BrClNO6
Formula Weight: 408.63Product specifications:Form: White to off-white crystalline powderAssay (TLC): ≥ 99%Chromatography (TLC): Homogeneous Identity: By IRSolubility (1%, MeOH): Clear and colorless[α]25
D : +145.0 ± 3.0° (c=0.5, DMF:H2O 1:1)Moisture (KF): ≤ 1.0%Storage: -20°C, protect from moisture.CAS #107021-38-5
26200 100 mg 500 mg
5-Bromo-4-Chloro-3-Indolyl-β-D-Galactopyranoside(X-GAL)
ULTRAPURE
C14H15BrClNO6
Formula Weight: 408.63Product specifications:Form: White crystalline powderAssay (HPLC): ≥ 99%Chromatography (TLC): HomogeneousIdentity: By IRSolubility (2%, DMF): Clear and colorlessApplications:� A histochemical substrate for β-Galactosidase� A substrate for blue/white colony screening.
Effective concentration is approximately 50 µg/ml.
Stock Concentration: 20 mg/ml in dimethylfor-mamide in a glass or polypropylene tube.
Storage: -20°C, protect from moisture.CAS #7240-90-6
10077 100 mg 250 mg 1 gm 2 gm 5 gm
5-Bromo-4-Chloro-3-5-Bromo-4-Chloro-3-Indolyl Phosphate, Disodium Salt(BCIP)
ULTRAPURE
C8H4BrClNNa2O4PFormula Weight: 370.43Product specifications:Form: White to off-white crystalline powderIdentity: By IRChromatography (TLC): HomogeneousStorage: -20°C, protect from moisture.CAS #102185-33-1
12387 100 mg 500 mg 1 gm
5-Bromo-4-Chloro-3-Indolyl Bromocresol Green, Sodium SaltULTRAPURE
ACS Reagent GradeC21H13Br4O5SNaFormula Weight: 720.00Product specifications:Form: Brown to orange powderClarity of Solution (0.1%, H2O): Passes testVisual Transition Interval: pH 3.8 - Yellow pH 5.4 - BlueApplication: � Used as a tracking dye in alkaline agarose gel
electrophoresis.Loading Buffer (6X): 6 mM EDTA 300 mM NaOH 18% Ficoll (in water) 0.15% Bromocresol Green 0.25% Xylene CyanolCAS #62625-32-5
12355 5 gm 25 gm
Biochemicals
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Bromophenol Blue, Sodium SaltULTRAPURE
ACS Reagent GradeC19H9Br4NaO5SFormula Weight: 691.94Product specifications:Form: PowderClarity of Solution: Passes testVisual Transition Interval: pH 3.0 - Yellow pH 4.6 - BlueApplications:� Used as a tracking dye in agarose and acrylamide
gel electrophoresis.� Used in the stop solution in DNA sequencing kits.CAS #62625-28-9
12370 5 gm 10 gm
BSA
See Albumin, Bovine, page 198
Buffers
Glycerol Tolerant Gel (GTG) Buffer, 20X Solution, page 219
HEPES, 1 M Solution, pH 7.3, page 222PBS, 10X Solution, pH 7.4, page 232RapidRun™ Agarose Buffer, 20X Solution,
page 238TAE Buffer, 10X Solution, page 243TAE Buffer, 50X Solution, page 243TBE Buffer, 5X Solution, page 243TBE Buffer, 10X Ready-Mixed Powder, page 243TE Buffer, 1X Solution, page 244TE Buffer, 1X Solution, pH 7.0, page 244TE Buffer, 1X Solution, pH 7.0, Low EDTA, page 244TE Buffer, 1X Solution, pH 8.0, page 244TE Buffer, 1X Solution, pH 8.0, Low EDTA, page 244TE Buffer, 50X Solution, page 244Tris-Glycine (TG) Buffer, 10X Solution, page 247Tris-Glycine-SDS (TGS) Buffer,10X Solution,
page 247Tris/HCl, 1 M Solution, pH 7.0, page 248Tris/HCl, 1 M Solution, pH 7.5, page 248Tris/HCl, 1 M Solution, pH 8.0, page 248
Cacodylic Acid, Sodium Salt Trihydrate(Dimethylarsenic Acid, Sodium Salt; Sodium Cacodylic)
(CH3)2AsO2Na•3H2OFormula Weight: 214.03 (trihydrate)Product specifications:Form: White powderAssay: ≥ 98.5% (dry basis)Solubility (10%, H2O): Clear and colorlessChloride (Cl): ≤ 0.1%Sulfate (SO4): ≤ 0.3%Application: � An integral component of terminal deoxynucleo-
tidyl transferase bufferNote: Material is hygroscopicCAS #124-65-2
12488 100 gm 500 gm
Caffeine, Anhydrous(1,3,7-Trimethylxanthine)
ULTRAPURE
C8H10N4O2
Formula Weight: 194.19Product specifications:Form: White powderAssay: 98.5 - 101.0%Identity (A, B): Passes testsMelting Point: 235 - 239°CLoss on Drying: ≤ 0.5%Residue on Ignition: ≤ 0.1%Heavy Metals: ≤ 10 ppmReadily Carbonizable Substances: Passes testOther Alkaloids: Passes testChromatographic Purity: Passes testResidual Solvents: Passes testCAS #58-08-2
12520 100 gm 1 kg
Calcium Chloride, Dihydrate
ACS Reagent GradeCaCl2•2H2OFormula Weight: 147.01Product specifications:Form: White crystalline powderAssay: 99.0 - 105.0%pH (5%, H2O, 25°C): 4.5 - 8.5Insoluble Matter: ≤ 0.01%Oxidizing Substances (NO3): ≤ 0.003%Sulfate (SO4): ≤ 0.01%Ammonium (NH4): ≤ 0.005%Barium (Ba): ≤ 0.005%Heavy Metals (Pb): ≤ 5 ppmIron (Fe): ≤ 0.001%Magnesium (Mg): ≤ 0.005%Potassium (K): ≤ 0.01%Sodium (Na): ≤ 0.02%Strontium (Sr): ≤ 0.1%CAS #10035-04-8
12531 500 gm 2.5 kg
D-Calcium Pantothenate(Pantothenic Acid, Calcium Salt; Vitamin B5 )
USBioAnalyzed(C9H16NO5)2CaFormula Weight: 476.53Product specifications:Form: White powderIdentity (A, B): Passes tests[α]25
D : +25.0° to +27.5° (c=5 in H2O, dry basis)Loss on Drying: ≤ 5.0%Heavy Metals (Pb): ≤ 20 ppmOrdinary Impurities: Passes testAlkalinity: Passes testNitrogen Content (N): 5.7 to 6.0%Calcium Content (Ca): 8.2 to 8.6%Residual Solvents: Passes testCAS #137-08-6
12550 100 gm 500 gm 1 kg
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Biochemicals
205For bulk or alternate pack sizes, email us at [email protected].
Carbenicillin, Disodium Salt
C17H16N2Na2O6SFormula Weight: 422.36Product specifications:Form: White to off-white crystalline powderAssay: 89.0 - 100.5% (dry basis)Identity: Passes testMoisture (KF): ≤ 5.0%Storage: 4°CCAS #4800-94-6
12678 1 gm 5 gm
Carboxypeptidase Y
(E.C.3.4.16.5)Source: YeastProduct specifications:Form: A useful enzyme in the sequence analysis of
proteins and peptides.Specific Activity: ≥ 100 units/mg proteinUnit Definition: One unit hydrolyzes 1 µmol of CBZ-
Phe-Leu/minute at 25°C, pH 6.5.Solubility: Soluble in water and phosphate bufferNote: Please inquire for bulk pricing.Storage: -20°C
12758 1 mg 10 mg
Casein, HammarstenULTRAPURE
Formula Weight: 75,000 - 100,000Product specifications:Form: Cream to light yellow, lyophilized powderProtein: ≥ 95%Loss on Drying: ≤ 10.0%CAS #9000-71-9
12840 100 gm 1 lb 5 lb 25 lb
Casein (High Nitrogen)
Prepared from New Zealand casein exclusivelyProduct specifications:Form: White to off-white powderProtein (dry basis): ≥ 95.0%CAS #9000-71-9
12845 5 lb 25 lb 100 lb
Casein Hydrolysate, Enzymatic DigestULTRAPURE
Product specifications:Form: PowderMoisture: ≤ 6%pH: 6.6 - 7.5Amino Nitrogen: 3.7 - 4.8%Total Nitrogen: 11.5 - 14.2%Protein (N x 6.38): 73.3 - 90.6%Ash: ≤ 6.5%CAS #65072-00-6
12855 1 lb 5 lb 25 lb
Casein, Sodium(Sodium Caseinate)
Product specifications:Form: White to off-white powderProtein (dry basis): ≥ 93%Ash: ≤ 4.2%Moisture: ≤ 5.0%pH (5%, H2O): 6.4 - 7.0CAS #9005-46-3
12865 1 lb 5 lb
Casein - Vitafree™ULTRAPURE
Product specifications:Form: Cream colored powderProtein (N x 6.25): ≥ 95.0%Loss on Drying: ≤ 12.0%Specifically prepared as a source of high quality
protein in diets needed to produce vitamin defi-ciencies.
CAS #68308-23-6
12866 5 lb 25 lb
Catalase
(E.C.1.11.1.6)Source: Bovine LiverProduct specifications:Form: Lyophilized powder, approx. one-third sucroseIdentity: By IRActivity: ≥ 3000 units/mg proteinProtein Concentration: Approx. 50%Unit Definition: One unit decomposes 1 µmol of
H2O2/minute at 25°C, pH 7.0.Storage: 4°C
12885 500 mg 1 gm 5 gm 10 gm
Cellulase
(E.C.3.2.1.4)Source: Aspergillus nigerUnit Definition: One unit produces a change in the
relative fluidity in a defined sodium carboxy-methyl cellulose substrate of 1 in 5 minutes at 40°C, pH 4.5.
Storage: 4°C
13285 100 gm
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Biochemicals
206 888-362-2447 | 216-765-5000 | usb.affymetrix.com
Cesium ChlorideULTRAPURE
MB GradeCsClFormula Weight: 168.36Product specifications:Form: White/clear crystalline powderAssay: 99.999%Refractive Index (50% w/w, 20°C): 1.389 - 1.391A (260 nm, 50% w/v, 1 cm): ≤ 0.02Insoluble Matter: NegligibleSilicon (Si): ≤ 1 ppmSulfate (SO4): ≤ 5 ppmBarium (Ba): ≤ 2 ppmCalcium (Ca): ≤ 3 ppmLithium (Li): ≤ 1 ppmMagnesium (Mg): ≤ 1 ppmPotassium (K): ≤ 3 ppmRubidium (Rb): ≤ 4 ppmSodium (Na): ≤ 3 ppmStrontium (Sr): ≤ 1 ppmAluminum (Al): ≤ 1 ppmChromium (Cr): ≤ 1 ppmIron (Fe): ≤ 5 ppmLead (Pb): ≤ 2 ppmManganese (Mn): ≤ 1 ppmDNase (endo): Passes testDNase (exo): Passes testRNase: Passes testApplication:� Used for density gradient high speed centrifuga-
tion separation of λ phage, M13 and plasmid DNA.
Densities in CsCl: Double-stranded DNA: density (P) = (0.098)
[G + C] + 1.660 gm/cm3 mole Sample Density RFI (Supercoiled) ds DNA 1.709 gm/cm3
RFII (Nicked) ds DNA 1.54 gm/cm3
ss DNA 1.726 gm/cm3
ss RNA 1.90 gm/cm3
Protein 1.33 gm/cm3
Note:1. Nicked DNA binds more ethidium bromide than
supercoiled DNA.2. Ethidium bromide at saturation decreases density
of ds DNA ~ 0.15 gm/cm3.3. More ethidium bromide bound = greater reduc-
tion in density.Typical Cesium Chloride Solutions: Step gradients prepared in SM* or H2O; for
(100 ml) Density CsCl SM or Refractive (P) (gm) H2O (ml) Index
1.45 60 85 1.377 1.50 67 82 1.382 1.70 75 75 1.399* SM—phage dilution buffer (100 mM NaCl, 8 mM MgSO4
•7H2O, 50 mM Tris-HCl, pH 7.5, 0.01% gelatin).
Cesium Chloride RNA Isolation Method:A. Guanidine Thiocyanate Homogenization Buffer
for RNA Isolation: 4.0 M Guanidine Thiocyanate, 0.1 M Tris, pH 7.5, 1.0% 2-mercaptoethanol
B. Cesium Chloride RNA Isolation Solution: Most researchers layer their RNA homogenates onto a cushion of 5.7 M Cesium Chloride.
C. RNA Resuspension Buffer: TE, pH 7.5.Reference:1. Schildkraut, C. L., Marmur, J. and Doty, P. (1962)
J. Mol. Biol. 4, 430-443.CAS #7647-17-8
75822 5 gm 25 gm 100 gm 500 gm 1 kg
Cetyl Trimethyl Ammonium Bromide(CTAB)
[CH3(CH2)15N(CH3)3]BrFormula Weight: 364.45Product specifications:Form: White crystalline powderAssay: ≥ 98.0% as isIdentity: By IRpH (10%, H2O): 3.0 - 7.0Moisture (KF): ≤ 1.0%Melting Point: 232 - 237°CApplications:� Cationic detergent, soluble in H2O and readily
soluble in alcohol.� Used in the preparation and purification of
genomic DNA from bacteria.� Complexes with both polysaccharide and residual
protein.� Also used for DNA mini preps for sequencing.CAS #57-09-0
13353 100 gm 500 gm 1 kg
Chicken Intestine Alkaline Phosphastase
E.C.3.1.3.1Source: Chicken IntestineProduct specifications:Form: Lyophilized powder salt freeUnit Definition: One unit hydrolyzes one µmole of
o-carboxyphenyl phosphate/minute at 25°C, pH 8.8.
Storage:-20°C, desiccated CAS #9001-78-9
10925 1 gm 5 gm
ChloramphenicolULTRAPURE
USBioAnalyzedC11H12Cl2N2O5
Formula Weight: 323.13Product specifications:Form: White to grey-white crystalline powderAssay: 97.0 - 103.0% (as is)Identity (A, B): Passes testsChromatographic Purity: Passes testMelting Point: 149 - 153°CpH (2.5%, H2O): 4.5 - 7.5Crystallinity: Passes testApplications:� Used in large-scale plasmid DNA preparation
of low to moderate copy number plasmids. Also used in high copy plasmids to limit the bulk and viscosity of the cell lysates
� Used in chloramphenicol acetyl transferase (CAT) assay
Mode of Action: Binds to the ribosomal 50S subunit and inhibits
protein synthesis. Soluble in methanol, ethanol.Stock Solution: 34 mg/ml in ethanol for plasmid
DNAsWorking Concentration: 25-170 µg/ml; 100 mg/ml
in ethanol for CAT assaysCAS #56-75-7
23660 25 gm 100 gm 1 kg
Cholesterol
C27H46OFormula Weight: 386.65Product specifications:Form: White to faintly yellow crystalline powderIdentity: Passes testSolubility in alcohol (1%): Passes test[α]20
D : -34° to -38° (c=2 in dioxane)Acidity: Passes testMelting Range: 147 - 150°CLoss on Drying: ≤ 0.3%Residue on Ignition: ≤ 0.1%CAS #57-88-5
13610 100 gm 1 kg
Cholesterol
Reagent GradeC27H46OFormula Weight: 386.65Storage: 4°CCAS #57-88-5
13580 5 gm 100 gm
Biochemicals
207For bulk or alternate pack sizes, email us at [email protected].
Choline Chloride
C5H14ClNOFormula Weight: 139.62Product specifications:Form: White crystalline powderAssay: ≥ 98.0% (dry basis)Identity: By IRMoisture (KF): ≤ 0.5%Residue on Ignition: ≤ 0.05%Heavy Metals (Pb): ≤ 10 ppm1,4-Dioxane: Passes testProtect from moisture. This material is hygroscopic.CAS #67-48-1
13410 200 gm 1 kg 5 lb 25 lb
Citric Acid, Anhydrous
USBioAnalyzedC6H8O7
Formula Weight: 192.12Product specifications:Form: White crystalline powderAssay: 99.5 - 100.5% (dry basis)Clarity of Solution: Passes testColor of Solution: Passes testIdentity: By IRMoisture (KF): ≤ 1.0%Residue on Ignition: ≤ 0.1%Readily Carbonizable Substances: Passes testHeavy Metals (Pb): ≤ 0.001%Oxalate: Passes testSulfate (SO4): Passes testCAS #77-92-9
13729 500 gm 1 kg
Citric Acid, Trisodium Salt, Dihydrate(Citric Acid, Sodium Salt; Sodium Citrate, Dihydrate)
ULTRAPURE
USBioAnalyzedC6H5Na3O7
•2H2OFormula Weight: 294.10Product specifications:Form: White crystalline powderAssay: 99.0 - 100.5% (dry basis)Identity A: By IRIdentity B: Passes testLoss on Drying: 10.0 - 13.0%Tartrate: Passes testAlkalinity: Passes testHeavy Metals (Pb): ≤ 10 ppmCAS #6132-04-3
13735 1 kg 5 kg
Cocarboxylase(Thiamine Pyrophosphate Chloride)
C12H19CIN4O7P2SFormula Weight.: 460.77Product specifications:Form: White powderAssay: ≥ 93.0% (dry basis)Identity: By IRLoss on Drying (130°C): ≤ 1.5%Chloride (Cl): 7.5 - 7.9%Sulfate (SO4): ≤ 0.01%Heavy Metals (Pb): ≤ 10 ppmCAS #154-87-0
13765 25 gm 100 gm
Coenzyme A, Free Acid, Trihydrate
C21H36N7O16P3S•3H2OFormula Weight: 821.58Product specifications:Form: Lyophilized powderAssay: ≥ 75% as reduced Co-A•3H2OSpectral Ratios (pH 7.5): A250/A260: 0.78 ± 0.03 A280/A260: 0.16 ± 0.03Moisture (KF): ≤ 8%Note: The above product is available in manufactur-
ing quantities. Please inquire for bulk pricing.Storage: -20°CCAS #85-61-0
13787 50 mg 100 mg 250 mg 500 mg 1 gm
Coenzyme A, Trilithium Salt, Dihydrate
C21H33N7O16P3SLi3•2H2OFormula Weight: 821.36Product specifications:Form: White crystalline powderAssay (Enzymatic): ≥ 75%Spectral Ratios (pH 7.5): A250/A260: 0.78 ± 0.03 A280/A260: 0.16 ± 0.03Lithium (Li): 2.5 ± 1.5%Moisture (KF): ≤ 8%Note: The above product is available in manufactur-
ing quantities. Please inquire for bulk pricing.Storage: -20°C, protect from moisture.CAS #18439-24-2
13785 100 mg 250 mg 500 mg 1 gm
Coenzyme I
See β-Nicotinamide Adenine Dinucleotide, page 229
Coenzyme II
See Nicotinamide Adenine Dinucleotide Phosphate, page 230
Collagen
Source: Calf SkinProduct specifications:Form: Solution in dilute (0.075 M) acetic acid with
0.01% thimerosal as a preservative, pH 3.7.Collagen concentration: Approximately 10 mg/mlStorage: 4°C
13813 10 ml 25 ml 100 ml
Collagenase, Type I
(E.C.3.4.24.3)Source: Clostridium histolyticumDescription: Contains a mixture of collagenase,
clostripain, trypsin, caseinase, and other lytic enzymes.
Product specifications:Form: Lyophilized powderActivity: ≥ 125 units/mg powder (dry basis) ≥ 0.15 Wunsch units/mg powder (dry basis)Unit Definition: One unit liberates 1 µmol of
L-Leucine equivalents from collagen in 5 hours at 37°C, pH 7.5. One unit is equal to 0.0012 Wunsch units.
Contaminating Activities: Caseinase: 200 - 1,000 units/mg Clostripain: 1.0 - 3.5 units/mg Trypsin: 0.1 - 0.5 units/mgStorage: 4°C, desiccatedCAS #9001-12-1
13820 100 mg 1 gm 5 gm
Compact Reaction Columns(CRC)
With screw-on lid, Luer-lock lid, stopper and filter insertion tools.
Upper and lower filters with a pore size appropri-ate to the customer’s needs must be purchased separately.
50 columns/pk
13928 1 pk
Biochemicals
208 888-362-2447 | 216-765-5000 | usb.affymetrix.com
Compact Reaction Columns(CRC) without screw on lids
Stopper and filter insertion tools included.Upper and lower filters with a pore size appropriateto the customer’s needs must be purchasedseparately.100 columns/pk
13929 1 pk
CRC Lower Filters(2.7 mm)
10 μm pore size50 filters/pk
13911 1 pk
CRC Lower Filters(2.7 mm)
35 μm pore size50 filters/pk
13912 1 pk
CRC Lower Filters(2.7 mm)
90 μm pore size50 filters/pk
13913 1 pk
CRC Upper Filters(6.8 mm)
10 μm pore size50 filters/pk
13914 1 pk
CRC Upper Filters(6.8 mm)
35 μm pore size50 filters/pk
13918 1 pk
CRC Upper Filters(6.8 mm)
90 µm pore size50 filters/pk
13919 1 pk
Coomassie Stains
See Brilliant Blue G & R, page 203
Creatine Kinase
(E.C.2.7.3.2)Source: Rabbit MuscleForm: Salt-free lyophilized powderUnit Definition: One unit phosphorylates 1 µmol of
creatine/minute at 25°C, pH 9.0.Storage: -20°C, desiccatedCAS #9001-15-4
13915 100 mg 1 gm
Creatine Phosphate, Disodium Salt, Tetrahydrate(Phosphocreatine)
C4H8N3O5PNa2•4H2O
Formula Weight: 327.14Product specifications:Form: White crystalline powderAssay: ≥ 98.0%Chromatography (TLC): HomogeneousIdentity: By IRMoisture (KF): 23.5 ± 2.5%Storage: 4°CCAS #922-32-7
13920 5 gm 25 gm
CTAB
See Cetyl Trimethyl Ammonium Bromide, page 206
CTP
See Cytidine-5’-Triphosphate, page 209
Cyanocobalamin
See Vitamin B12, page 250
α-Cyclodextrin
C36H60O30
Formula Weight: 972.84Product specifications:Form: White crystalline powderAssay (HPLC): ≥ 98.0%[α]20
D : +147° to +152° (c=1 in H2O)Loss on Drying: ≤ 12%Residue on Ignition: ≤ 0.1%Chloride (Cl): ≤ 0.018%Arsenic (As): ≤ 2 ppmHeavy Metals (Pb): ≤ 10 ppmCAS #10016-20-3
13979 25 gm
L-Cysteine, Hydrochloride, Monohydrate
HSCH2CH(NH2)COOH•HCl•H2OFormula Weight: 175.63Product specifications:Form: White crystalline powderAssay: 98.5 - 101.5% (dry basis)Identity: By IRChromatography: Homogeneous[α]25
D : +5.7° to +6.8° (c=8 in in 6N HCl, dry basis)Loss on Drying: 8.0 - 12.0%Residue on Ignition: ≤ 0.1%Sulfate (SO4): ≤ 0.03%Heavy Metals (Pb): ≤ 10 ppmArsenic (As): ≤ 1.5 ppmChloride (Cl): 19.8 - 20.8%Iron (Fe): ≤ 10 ppmCAS #7048-04-6
14035 25 gm 100 gm 1 kg 5 kg
L-Cystine
CP GradeC6H12N2O4S2
Formula Weight: 240.30Product specifications:Form: White crystalline powderAssay: 98.5 - 101.5% (dry basis)Identity: By IR[α]20
D : -215° to -225° (c=2 in 1N HCl)Loss on Drying: ≤ 0.2%Residue on Ignition: ≤ 0.1%Heavy Metals (Pb): ≤ 20 ppmLead (Pb): ≤ 10 ppmCAS #56-89-3
14050 50 gm 100 gm 1 kg 5 kg
Cytidine, Free Base
C9H13N3O5
Formula Weight: 243.22Product specifications:Form: White crystalline powderAssay (UV): ≥ 97.5%Identity: By IRλmax (pH 1.0): 280 nm ± 2 nmSpectral Ratios (pH 1.0): A250/A260: 0.45 ± 0.02 A280/A260: 2.10 ± 0.05Loss on Drying: ≤ 1.0%Reference constants:λmax (pH 7, NRC): 271 nmεmax (pH 7, NRC): 9.1 x 103
ε260 (pH 7, DBR): 7.55 x 103
CAS #65-46-3
14060 25 gm 100 gm
Biochemicals
209For bulk or alternate pack sizes, email us at [email protected].
Cytidine-5’-Triphosphate, Disodium Salt, Dihydrate(CTP)
C9H14N3O14P3Na2•2H2O
Formula Weight: 527.13 (anhydrous)Formula Weight: 563.16 (dihydrate)Product specifications:Form: White powderAssay (Total UV, pH 2.0): ≥ 96.0%λmax: 280 nm ± 1 nmεmax: (pH 2.0): [13.0 ± 0.52] x 103
Spectral Ratios (pH 2.0): A250/A260: 0.45 ± 0.05 A280/A260: 2.10 ± 0.07Moisture (KF): ≤ 12.0%Reference constants:λmax (pH 7, NRC): 271 nmεmax (pH 7, NRC): 9.1 x 103
ε260 (pH 7, DBR): 7.4 x 103
Storage: -20°C, protect from moisture.CAS #81012-87-5
14121 100 mg 500 mg 1 gm
Cytochrome C
Source: Horse HeartProduct specifications:Form: Reddish or dark brown crystalline powderAssay: ≥ 90.0% (dry basis)Identity: Passes testpH (1%, H2O): 5.0 - 7.0Moisture (KF): ≤ 6.0%Iron (Fe): 0.40 to 0.50%Residue on Ignition: ≤ 1.5%Solubility (10%, H2O): Clear dark red solutionStorage: -20°CCAS #9007-43-6
14102 100 mg 500 mg 1 gm
DAPI(4’,6-Diamidino-2-Phenylindole Dihydrochloride)
C16H15N5•2HCl
Formula Weight: 350.25Product specifications:Form: Yellow powderAssay (HPLC): ≥ 98%Identity: By IRSolubility (1%, H2O): ClearNitrogen: ≥ 18.0%Applications:� Cytochemical technique for demonstration of
DNA in cells infected with mycoplasmas and viruses(1)
� Interaction of DAPI with synthetic polynucleo-tides(2)
� DAPI-stained cells can be fixed for high resolution flow cytometry(3).
� Reverse fluorescent chromosome banding by double-staining with chromomycin and DAPI(4)
� Binding of DAPI to GC and mixed sequences in DNA with an intercalation of a groove-binding molecule(5)
Notes: DAPI is an excellent dye for the staining of DNA. Originally, only the specific binding to AT-base pairs without an intercalation was known(2). Later on, the intercalation into GC-base pairs was shown. The most popular application of DAPI is its use as a reagent to detect mycoplasma or virus DNA (e.g. vaccinia infection or ‘unwant-ed’ viral contamination of cell culture cells) in cell culture. Reference 1 describes a simple procedure to stain DNA with DAPI: Grow cells on a coverslip in a cell culture dish. Wash the coverslip once with PBS. Incubate the cells on the coverslip at 37°C for 15 - 30 minutes in 0.1 µg DAPI/ml PBS. Wash the coverslip in PBS and put it up-side-down on a slide. Examine the cells under a micro-scope (excitation: 365 nm; emission maximum at 450 nm). Prolonged incubation with DAPI increases the nuclear fluorescence; shorter incu-bation time leads to a weaker nuclear staining, which facilitates the examination of the cytoplas-mic fluorescence.
DAPI can be dissolved in water, PBS, 5 mM Tris-HCl (pH 7.6)/8 mM NaCl, citrate or phosphate buf-fer(3). The solubility is approximately 2.5%. Stock solutions are prepared at concentrations ranging from 1-2 mg/ml and the working dilution is 1:1000. DAPI solution is stable at room tempera-ture for 1 - 2 weeks(3), at 4°C up to 6 months, and at -20°C for 6 - 12 months. DAPI has hydrolyzed if the solution becomes turbid. DAPI bleaches quickly in contact with light, even though it is quite stable against UV-light.
References:1. Russel, W. C., Newman, C. and Williamson, D. H.
(1975) Nature 253, 461-462.2. Kapuscinski, J. and Szer, W. (1979) Nucleic Acids
Res. 6, 3519-3534. 3. Otto, F. (1990) Methods Cell Biol. 33, 105-110.4. Schweizer, D. (1976) Chromosoma. 29, 307-324.5. Wilson, W. D., Tanious, F. A., Barton, H. J.,
Jones, R. L., Fox, K., Wydra, R. L. and Strekowski, L. (1990) Biochem. 11, 8452-8461.
Storage: 4°CCAS #28718-90-3
14564 10 mg 50 mg
Denhardt’s Solution, 50XULTRAPURE
MB GradeForm: 50X solution of 1% acetylated bovine serum
albumin (0.2 µm filtered), 1% Ficoll (autoclaved before use), and 1% PVP-90 (autoclaved before use). Dispensed into autoclaved bottles.
Contaminant Testing: DNase (endo): Passes test DNase (exo): Passes test RNase: Passes test Protease: Passes testStorage: -20°CApplication: � For use as a blocking reagent in nucleic acid
hybridization techniquesFormamide Hybridization Buffer: 50% Formamide,
5X SSC, 0.5% SDS, 5X Denhardt’s Solution, 100 µg/ml sonicated fish sperm DNA
Reagent Amount for every 100 mlFormamide* 50 ml20X SSC* 25 ml20% SDS* 2.5 ml50X Denhardt’s Solution* 10 ml10 mg/ml fish DNA 1.0 mlWater* up to 100 ml*Denotes Ultrapure Reagent available.Reference:1. Sambrook, J., Fritsch, E. F. and Maniatis, T. (1989)
Molecular Cloning, 2nd Edition, Cold Spring Harbor Laboratory.
70468 50 ml
2’-Deoxyadenosine
C10H13N5O3•H2O
Formula Weight: 269.26Product specifications:Form: White to off-white crystalline powderAssay (Total UV): ≥ 98%Identity: By IRλmax: 258-260 nmSpectral Ratios: A250/A260: 0.77 ± 0.04 A280/A260: 0.15 ± 0.05Moisture (KF): ≤ 8.0%Reference constants:λmax (pH 7, NRC): 260 nmεmax (pH 7, NRC): 14.9 x 103
ε260 (pH 7, DBR): 15.2 x 103
CAS #16373-93-6
14235 5 gm 25 gm
Biochemicals
210 888-362-2447 | 216-765-5000 | usb.affymetrix.com
2’-Deoxyadenosine-5’-Triphosphate, Sodium Salt, 100 mM Solution(dATP)
ULTRAPURE
C10H12N5O12P3Na4
Formula Weight: 579.11Product specifications:Form: 100 mM aqueous solutionAssay (HPLC): ≥ 99%pH: 7.5 ± 0.2λ max: 259 nm ± 1 nmSpectral Ratios: A250/A260: 0.80 ± 0.03 A280/A260: 0.12 ± 0.02Functional Tests: Functionally tested in long PCR to
generate a 20.7 kb fragment.Storage: -20°C
77102 25 µmol (250 µl) 100 µmol (1 ml) 500 µmol (5 ml)
Deoxycholic Acid, Sodium SaltSodium Deoxycholate
C24H39NaO4
Formula Weight: 414.55Product specifications:Form: White to off-white powderAssay (HPLC): ≥ 98% (dry basis)Identity: By IRLoss on Drying (4 hours, 140°C, vacuum): ≤ 6%Sodium Cholate: ≤ 2%CAS #302-95-4
21026 25 gm 100 gm
2’-Deoxycytidine, Hydrochloride, Synthetic
C9H13N3O4•HCI
Formula Weight: 263.68Product specifications:Form: White crytalline powderAssay (Total UV): ≥ 98% (dry basis)Identity: By IRλmax: 280 ± 2 nmεmax (pH 1.0): 13.2 x 103 ± 2%Spectral Ratios (pH 1.0): A250/A260: 0.43 ± 0.03 A280/A260: 2.10 ± 0.05Moisture (KF): ≤ 1%Heavy Metals: ≤ 10 ppmCAS #3992-42-5
14284 1 gm 5 gm 10 gm 100 gm
2’-Deoxycytidine-5’-Triphosphate, Sodium Salt, 100 mM Solution(dCTP)
ULTRAPURE
C9H12N3O13P3Na4
Formula Weight: 555.08Product specifications:Form: 100 mM aqueous solutionAssay (HPLC): ≥ 99%pH: 7.5 ± 0.2λ max (pH 2.0): 280 nm ± 1 nmSpectral Ratios (pH 2.0): A250/A260: 0.45 ± 0.03 A280/A260: 2.10 ± 0.15 A290/A260: 1.60 ± 0.10Functional Tests: Functionally tested in long PCR to
generate a 20.7 kb fragment.Storage: -20°C
77104 25 µmol (250 µl) 100 µmol (1 ml) 500 µmol (5 ml)
2’-Deoxyguanosine
C10H13N5O4•H2O
Formula Weight: 285.26Product specifications:Form: White granular powderAssay (HPLC): ≥ 97%Identity: By IRChromatography (TLC): Homogeneousλmax: 253 ± 2 nmεmax: 12800-14300Spectral Ratios (pH 7.0): A250/A260: 1.09 - 1.21 A280/A260: 0.63 - 0.71Moisture (KF): ≤ 9%Reference constants:λmax (pH 7, NRC): 254 nmεmax (pH 7, NRC): 13.0 x 103
CAS #961-07-9
14300 5 gm 25 gm 100 gm
2’-Deoxyguanosine, Monohydrate Synthetic
C10H13N5O4•H2O
Formula Weight: 285.26Product specifications:Form: White crystalline powderAssay: ≥ 98% (dry basis)Chromatography (TLC): HomogeneousIdentity: By IRλmax (pH 7.0): 253 ± 2 nmεmax (pH 7.0): (13.7 x 103) ± 4%Spectral Ratios (pH 7): A250/A260: 1.12 - 1.18 A280/A260: 0.65 - 0.69Moisture (KF): ≤ 7.0%CAS #961-07-9
14304 1 gm 5 gm 10 gm 100 gm 1 kg
7-deaza-2’-Deoxyguanosine-5’-Triphosphate, Lithium Salt, 10 mM Solution(7-deaza-dGTP)
C11H15N4O13P3Li2Formula Weight: 518.10Product specifications:Form: 10 mM aqueous solutionAssay (HPLC): ≥ 95%pH: 7.0λmax (pH 7.5): 259 nm ± 1 nmFunctional Tests: Meets or exceeds criteria for high
quality sequencing.Storage: -20°C
70065 2 µmol
2’-Deoxyguanosine-5’-Triphosphate, Sodium Salt, 100 mM Solution(dGTP)
ULTRAPURE
C10H12N5O13P3Na4
Formula Weight: 595.11Product specifications:Form: 100 mM aqueous solutionAssay (HPLC): ≥ 99%pH: 7.5 ± 0.2λmax: 252 nm ± 1 nmSpectral Ratios (pH 7.5): A250/A260: 1.18 ± 0.04 A280/A260: 0.67 ± 0.03 A290/A260: 0.28 ± 0.03Functional Tests: Functionally tested in long PCR to
generate a 20.7 kb fragment.Storage: -20°C
77106 25 µmol (250 µl) 100 µmol (1 ml) 500 µmol (5 ml)
Biochemicals
211For bulk or alternate pack sizes, email us at [email protected].
2’-Deoxynucleoside-5’-Triphosphates, Sodium Salt, 100 mM Solutions, Sets of Four(dNTPs)
ULTRAPURE
Form: 100 mM aqueous solutions, pH 7.5; dATP, dCTP, dGTP, dTTP
Functional Tests: Functionally tested in long PCR to generate a 20.7 kb fragment.
Storage: -20°C
4 x 25 µmol (250 µl) each dNTP per pack77100 1 pk4 x 100 µmol (1 ml) each dNTP per pack77328 1 pk4 x 500 µmol (5 ml) each dNTP per pack77128 1 pk
2’-Deoxynucleoside-5’-Triphosphate Mixes
See PCR Nucleotide Mixes, page 232
2-Deoxy-D-RiboseULTRAPURE
C5H10O4
Formula Weight: 134.13Product specifications:Form: White powderAssay: ≥ 98.0%[α]20
D : -57.3° ± 1.5°(c=1 in H2O)Loss on Drying: ≤ 0.5%Residue on Ignition: ≤ 0.5%Heavy Metals (Pb): ≤ 5 ppmIron (Fe): ≤ 5 ppmCAS #533-67-5
14420 5 gm 25 gm 100 gm
2’-Deoxythymidine-5’-Triphosphate, Sodium Salt, 100 mM Solution(dTTP)
ULTRAPURE
C10H13N2O14P3Na4
Formula Weight: 570.10Product specifications:Form: 100 mM aqueous solutionAssay (HPLC): ≥ 99%pH: 7.5 ± 0.2λmax: 267 nm ± 1 nmSpectral Ratios (pH 7.5): A250/A260: 0.65 ± 0.03 A280/A260: 0.73 ± 0.03 A290/A260: 0.28 ± 0.03Functional Tests: Functionally tested in long PCR to
generate a 20.7 kb fragment.Storage: -20°C
77108 25 µmol (250 µl) 100 µmol (1 ml) 500 µmol (5 ml)
2’-Deoxyuridine-5’-Triphosphate, Sodium Salt, 100 mM Solution(dUTP)
ULTRAPURE
C9H11N5O12P3Na4
Formula Weight: 556.1Product specifications:Form: 100 mM aqueous solutionAssay (HPLC): ≥ 99%pH: 7.5 ± 0.2λmax (pH 7.5): 262 nm ± 1 nmAbsorbance ratios: A250/A260: 0.74 ± 0.03 A280/A260: 0.35 ± 0.03Functional Test: Functionally tested in PCRStorage: -20°C
77206 25 µmol (250 µl) 100 µmol (1 ml) 500 µmol (5 ml)
DEPC
See Diethyl Pyrocarbonate, page 212
DextranULTRAPURE
M.W.: 60,000 - 90,000Product specifications:Form: White powderpH (6% w/v in KCl): 4.5 - 7.0Loss on Drying: ≤ 7.0%Residue on Ignition: ≤ 1.9%CAS #9004-54-0
14495 100 gm 1 kg
Dextran Sulfate, Sodium Salt
M.W.: 40,000 - 50,000Product specifications:Form: Off white to light yellow powderLoss on Drying: ≤ 10.0%pH (1%, H2O): 5.0 - 7.5Specific Viscosity (25°C in 1.0 M NaCl): 0.05 - 0.13Sulfur: 17.0 - 20.0%Glucose: 35.0 - 48.0%Residue on Ignition: 35.0 - 50.0%CAS #9011-18-1
14489 10 gm 50 gm 100 gm
Dextran Sulfate, Sodium SaltULTRAPURE
MW: 500,000*Product specifications:Form: White to off-white powderLoss on Drying: ≤ 10%pH (1%, H2O): 5.0-7.5Residue on Ignition: 40 - 50%* Note: Approximate molecular weight of dextran used in the production of the sulfate. Average MW of dextran sulfate sodium salt is approx. 1,400,000.
CAS #9011-18-1
70796 10 gm 50 gm
D-(+)-Dextrose, Anhydrous(α-D-(+)-Glucose)
CH2OH(CHOH)4CHOFormula Weight: 180.16Product specifications:Form: White crystalline powderIdentity: By IR[α]25
D : +52.6° to +53.2° (dry basis, c=10 in 0.012N NH4OH)
Moisture (KF): ≤ 1.0%CAS #50-99-7
14535 5 lb 25 lb 100 lb
Biochemicals
212 888-362-2447 | 216-765-5000 | usb.affymetrix.com
D-(+)-Dextrose, Monohydrate(α-D-(+)-Glucose)
C6H12O6•H2O
Formula Weight: 198.17Product specifications:Form: White crystalline powderIdentity: By IR[α]25
D : +52.6° to +53.2° (dry basis, c=10 in 0.12N NH4OH)
Moisture (KF): ≤ 9.5%Storage: Ambient, in tightly closed containers.CAS #50-99-7
14536 10 kg
Diaphorase
(E.C.1.8.1.4)Source: Clostridium kluyveriForm: Lyophilized powderUnit Definition: One DCPIP unit is that amount of enzyme which
causes a decrease in absorbance of 1.0 per min-ute at 25°C, pH 7.5, with 2,6-dichlorophenolin-odophenol as the chromogen.
One MTT unit is that amount of enzyme which reduces 1 µmol of MTT per minute at 30°C under stated assay conditions.
Activity (DCPIP units/mg): ≥ 30 DCPIP units/mg dry weight
Activity (MTT units/mg): ≥ 3.9 MTT units/mg dry weight
Storage: 4°C, desiccatedCAS #9001-18-7
14615 600 units 1,800 units
2’,3’-Dideoxyadenosine-5’-Triphosphate(ddATP)
ULTRAPURE
C10H12N5O11P3Na4
Formula Weight: 563.11Product specifications:Assay (FPLC): ≥ 99%pH: 7.5 ± 0.2λmax: 259 nm ± 1 nmSpectral Ratios (pH 7.5): A250/A260: 0.80 ± 0.03 A280/A260: 0.12 ± 0.02Functional Test: Meets or exceeds criteria for high
quality sequencing.Storage: -20°C
10 mM aqueous solution, pH 7.577110 0.5 µmol (50 µl) 1 µmol (100 µl)
2’,3’-Dideoxycytidine-5’-Triphosphate(ddCTP)
ULTRAPURE
C9H12N3O12P3Na4
Formula Weight: 539.08Product specifications:Assay (FPLC): ≥ 99%pH: 7.5 ± 0.2λmax (pH 2.0): 280 nm ± 1 nmSpectral Ratios (pH 2.0): A250/A260: 0.45 ± 0.03 A280/A260: 2.20 ± 0.20 A290/A260: 1.70 ± 0.20Functional Test: Meets or exceeds criteria for high
quality sequencing.Storage: -20°C
10 mM aqueous solution, pH 7.577112 0.5 µmol (50 µl) 1 µmol (100 µl)
2’,3’-Dideoxynucleoside-5’-Triphosphates, 10 mM Solutions, Sets of Four(ddNTPs)
ULTRAPURE
Form: 10 mM aqueous solutions, pH 7.5; ddATP, ddCTP, ddGTP, ddTTP
Storage: -20°C
4 x 0.5 µmol (50 µl) each ddNTP per pack77126 1 pk4 x 1.0 µmol (100 µl) each ddNTP per pack77125 1 pk
2’,3’-Dideoxynucleoside-5’-Triphosphates, 100 mM Solutions, Set of Four(ddNTPs)
ULTRAPURE
Form: 100 mM aqueous solutions, pH 7.5; ddATP, ddCTP, ddGTP, ddTTP
Storage: -20°C
4 x 4 µmol (40 µl) each ddNTP per pack77335 1 pk
2’,3’-Dideoxythymidine-5’-Triphosphate(ddTTP)
ULTRAPURE
C10H13N2O13P3Na4
Formula Weight: 554.10Product specifications:Assay (FPLC): ≥ 99%pH: 7.5 ± 0.2λmax (pH 7.5): 267 nm ± 1 nmSpectral Ratios (pH 7.5): A250/A260: 0.65 ± 0.03 A280/A260: 0.83 ± 0.03Functional Tests: Meets or exceeds criteria for high
quality sequencing.Storage: -20°C
10 mM aqueous solution, pH 7.577116 0.5 µmol (50 µl) 1 µmol (100 µl)
Diethyl Pyrocarbonate(DEPC)
ULTRAPURE
O(COOC2H5)2
Formula Weight: 162.14Product specifications:Form: Clear, colorless liquidAssay: ≥ 97%Density (20°C): 1.121 ± 0.005Refractive Index (20°C): 1.398 ± 0.005Applications:� A strong, nonspecific inhibitor of ribonuclease� Reacts with the N1 of histidine residues in pro-
teins under mild conditions.� Used to cross-link and modify proteins by eth-
oxyformylation of NH2 and imidazole groups in proteins.
� Reacts with amines and thus cannot be used with Tris Buffer.
References:1. Tsurushiin, S., Hiramatsu, A., Inamasu, M. and
Yasunobu, K. T. (1975) Biochim. Biophys. Acta. 400, 451-460.
2. Meyer, S. E. and Cromartie, T. H. (1980) Biochemistry 19, 1874-1881.
3. Berger, S. L. (1975) Anal. Biochem. 67, 428-437.4. Leonard, N. J., Mcdonald, J. J., Henderson, R. E.
and Reichmann, M. E. (1971) Biochemistry 10, 3335-3342.
5. Roberts, M. F., Deems, R. A., Mincey, T. C. and Dennis, E. A. (1977) J. Biol. Chem. 252, 2405-2411.
Storage: -20°CCAS #1609-47-8
14710 25 ml 100 ml
Biochemicals
213For bulk or alternate pack sizes, email us at [email protected].
Dihydrostreptomycin Sulfate
USBioAnalyzed(C21H41N7O12)2
•3H2SO4
Formula Weight: 1461.42Product specifications:Form: White to off-white powderActivity: ≥ 650 µg of C21H41N7O12/mg (as is)Identity (A, B): Passes testspH (200 mg dihydrostreptomycin/ml): 4.5 - 7.0Loss on Drying: ≤ 5.0%Streptomycin: ≤ 3.0%CAS #5490-27-7
14765 100 gm
Dimedone(5,5-Dimethyl-1,3-Cyclohexanedione)
C8H12O2
Formula Weight: 140.18Product specifications:Form: White to off-white crystalline powderAssay: ≥ 99%Identity: By IRMelting Point: 145 - 151°CSolubility (5% EtOH): Passes testStorage: Ambient, in tightly closed containers.CAS #126-81-8
14850 100 gm 1 kg
p-Dimethylamino Benzaldehyde
(CH3)2NC6H4CHOFormula Weight: 149.19Product specifications:Form: White to off-white powderIdentity: By IRChromatography (TLC): HomogeneousMelting Point: 70 - 75°CCAS #100-10-7
14868 100 gm 500 gm
p-Dimethylamino Cinnamaldehyde
(CH3)2NC6H4CH:CHCHOFormula Weight: 175.23Product specifications:Form: Bright yellow/gold crystalline powderAssay: ≥ 98.0%Identity: By IRMelting Point: 137 - 142°CCAS #6203-18-5
14872 25 gm
Dithioerythritol(DTE)
ULTRAPURE
C4H10O2S2
Formula Weight: 154.25Form: White crystalline powderChromatography (TLC): HomogeneousAssay (I2): ≥ 99%Melting Point: ≥ 81°CA (283 nm, 0.02 M, H2O): ≤ 0.04Storage: -20°C, protect from moisture.CAS #6892-68-8
15385 5 gm 25 gm 100 gm
Dithiothreitol(DTT; Cleland’s Reagent)
ULTRAPURE
MB GradeC4H10O2S2
Formula Weight.: 154.25Product specifications:Form: White crystalline powderAssay (SH): ≥ 99.5%A (280 nm, 0.1 M, 1 cm): ≤ 0.08A (260 nm, 0.1 M, 1 cm): ≤ 0.40Melting Point: 40 - 43°COxidized DTT: ≤ 0.3%DNase: Passes testRNase: Passes testProtease: Passes testApplication:� Used as a protective agent for SH groups.Reference:1. Cleland,W. W. (1964) Biochemistry 3, 480-482.Storage: -20°CCAS #3483-12-3
15397 1 gm 5 gm 25 gm 100 gm
Dithiothreitol(DTT; Cleland’s Reagent)
C4H10O2S2
Formula Weight: 154.25Product specifications:Form: White crystalline powderAssay: ≥ 97%Identity: By IRMelting Point: ≥ 38°COxidized DTT: ≤ 1%Storage: -20°C, in tightly closed containers.CAS #3483-12-3
15395 5 gm 25 gm 100 gm
Dithiothreitol (DTT), 0.1 M SolutionULTRAPURE
MB GradeProduct specifications:Form: 0.1 M solution of DTT in high purity dH2O.
Solution is 0.2 µm filtered and dispensed into vials.
Contaminant Testing: DNase (endo): Passes test DNase (exo): Passes test RNase: Passes testStorage: -20°C
70726 150 µl 1 ml 5 ml
DNA, Calf ThymusDNA, E. coliDNA, Fish SpermDNA, Fish Sperm, Activated DNA-Cellulose, Double-Stranded(dsDNA-Cellulose)DNA-Cellulose, Single-Stranded(ssDNA-Cellulose)
See Molecular Biology Products & Kits section, pages 128–129
DNase I
(E.C.3.1.21.1)Source: Bovine pancreasForm: Lyophilized powderActivity: >90,000 Dornase units/mg >2,500 Kunitz units/mgUnit Definition: One Kunitz unit causes an increase
in absorbance of 0.001 per minutes per ml at 260 nm at 25°C, pH 5, using the assay of Kunitz(1). One Kunitz unit = 35 Dornase units.
Reference:1. Kunitz (1950) J. Gen. Physiol. 33, 349-363.Storage: 4°C
14340 10 mg 20 mg
Dodecyl Sulfate, Lithium Salt
See Lithium Dodecyl Sulfate, page 225
Dodecyl Sulfate, Sodium Salt
See Sodium Dodecyl Sulfate, page 240
Biochemicals
214 888-362-2447 | 216-765-5000 | usb.affymetrix.com
N-Dodecyl Trimethylammonium Bromide(DTAB)
C15H34BrNFormula Weight: 308.34Product specifications:Form: White powderMelting Point: 239 - 247°CCAS #1119-94-4
18218 10 gm 50 gm 250 gm
DPN
See Nicotinamide Adenine Dinucleotide, page 229
DTAB
See N-Dodecyl Trimethylammonium Bromide, page 214
DTE
See Dithioerythritol, page 213
DTT
See Dithiothreitol, page 213
EDTA
See Ethylenediamine Tetraacetic Acid, page 215
Egg White
See Albumin-Egg, page 199
EGTA
See Ethylene Glycol-o-o’-bis (2-aminoethyl)-N,N,N’,N’,-Tetraacetic Acid, page 215
Elastase
(E.C.3.4.21.11)Source: Porcine PancreasForm: Lyophilized, salt free powderUnit Definition: One unit solubilizes 1 mg of elas-
tin/20 minutes at 37°C, pH 8.8.Storage: 4°CCAS #9004-06-2
15475 5 mg 10 mg 20 mg 100 mg
Enolase
(E.C.4.2.1.11)Source: YeastProduct specifications:Form: 50% glycerol solutionSpecific Activity: ≥ 40 units/mg proteinContaminating Activities: Phosphoglyceromutase: ≤ 0.02% Pyruvate Kinase: ≤ 0.02%Glycerol: 50%K-PO4: 0.1 M ± 0.05 MpH: 7.0Unit Definition: One unit oxidizes 1 µmol of NADH/
minute at 25°C, pH 7.5.Storage: -20°C
15515 10,000 units
Ergocalciferol
USBioAnalyzedC28H44OFormula Weight: 396.65Product specifications:Form: White to light yellow powderIdentification (A, B, C, D,): Passes testAssay: 97.0 - 103.0% (as is)[α]25
D : +103° to +106° (c= 1.5 in EtOH)Melting Range: 115 - 119°CReducing Substances: Passes testStorage: 4°C, protected from light sealed under
nitrogen.
15562 5 gm 25 gm 100 gm
Esculin
C15H16O9
Formula Weight: 340.28 (anhydrous)Product specifications:Form: White to off-white powderAssay: 97.5 - 102.5%Identity: By IR[α]20
D : -84° to -87° (c=2 in Dioxane:H2O/1:1, dry basis)
Moisture (KF): 6.5 - 8.0%CAS #66778-17-4
15590 100 gm
Ethidium BromideULTRAPURE
C21H20BrN3
Formula Weight.: 394.31Product specifications:Form: Purple/maroon crystalline powderAssay: ≥ 98.0%Residue on Ignition: ≤ 0.2%Ammonium Bromide: ≤ 0.1%Moisture (KF): ≤ 10.0%Applications:� A fluorescent dye that binds to DNA by intercalat-
ing between the bases, causing the double helix to unwind.
� Used to detect both single-and double-stranded DNA and RNA.
� Used in DNA isolation procedures and in gel elec-trophoresis.
Stock Solution: 10 mg/ml 100 mg Ethidium Bromide 10 ml H2O Filter and store solution protected from light.Working Concentration: 0.5 µg/ml for gel staining.Note: The electrophoretic mobility of linear double-
stranded DNA is reduced by 15% in the presence of ethidium bromide.
CAS #1239-45-8
32813 1 gm 5 gm 25 gm
Ethidium Bromide DropsULTRAPURE
Form: Ethidium bromide is supplied in a convenient, ready-to-dispense dropper format for gel electro-phoresis applications. One drop contains 40 µl or 25 µg of ethidium bromide.
Concentration: 0.625 mg/ml in water.Testing: Functionally tested for use in gel electro-
phoresis.To Use: Simply add one drop for every 50 ml of aga-
rose and/or buffer for a working concentration of 0.5 µg/ml.
75816 5 ml
Biochemicals
215For bulk or alternate pack sizes, email us at [email protected].
Ethylenediaminetetraacetic Acid, (EDTA), 0.5 M SolutionULTRAPURE
MB GradeForm: 0.5 M solution of EDTA•Na2 in high purity
dH2O, pH 8.0. Solution is 0.2 µm filtered and dis-pensed into durable, square Nalgene bottles.
pH: 8.0 ± 0.5Contaminant Testing: DNase (endo): Passes test DNase (exo): Passes test RNase: Passes test Protease: Passes testApplication: � Chelating agent used in many enzyme buffers
and, at higher concentrations, as an enzyme inac-tivator
15694 100 ml 500 ml
Ethylenediaminetetraacetic Acid, (EDTA) Disodium Salt, DihydrateULTRAPURE
C10H14N2Na2O8•2H2O
Formula Weight: 372.24Product specifications:Form: White crystalline powderAssay: 99.0 - 101.0%A (260 nm, 0.1 M, 1 cm): ≤ 0.2A (280 nm, 0.1 M, 1 cm): ≤ 0.05Insoluble Matter: ≤ 0.005%pH (5%, H2O): 4.0 - 6.0Nitrotriacetic Acid: ≤ 0.1%Heavy Metals (Pb): < 0.005%Iron (Fe): ≤ 0.01%Arsenic (As): ≤ 1 ppmCalcium (Ca): Passes testSolubility (0.1 M, H2O): Completely clear and color-
lessContaminant Testing: DNase (endo): Passes test DNase (exo): Passes test RNase: Passes testApplication: � Chelating agent used in many enzyme buffers
and as an enzyme inactivator at high concentra-tions
Stock Solution (0.5 M): 186.1 gm EDTA H2O to 800 ml Titrate with approximately 20 gm of NaOH pellets
to pH 8.0. Bring solution to 1 liter with water and sterilize by autoclaving.
CAS #6381-92-6
15701 100 gm 500 gm 1 kg
Ethylenediaminetetraacetic Acid, (EDTA) Disodium Salt, Dihydrate
ACS Reagent GradeC10H14N2O8Na2O8
•2H2OFormula Weight: 372.24Product specifications:Form: White crystalline powderAssay: 99.0 - 101.0%pH (5%, H2O, 25°C): 4.0 - 6.0Insoluble Matter: ≤ 0.005%Nitrilotriacetic Acid: ≤ 0.1%Heavy Metals (Pb): ≤ 0.005%Iron (Fe): ≤ 0.01%A (260 nm, 0.1 M, 1 cm): ≤ 0.2A (280 nm, 0.1 M, 1 cm): ≤ 0.05Solubility: (0.1 M, H2O): Clear and colorlessCAS #6381-92-6
15699 500 gm 1 kg 5 kg
Ethylenediaminetetraacetic Acid, (EDTA) Disodium Salt, Dihydrate
C10H14N2O8Na2•2H2O
Formula Weight: 372.24Product specifications:Form: White crystalline powderAssay: 99.0 - 101.0% (as disodium dihydrate)Identity: By IRSolubility (5%, H2O): Clear, colorless to slightly yel-
lowpH (5%, H2O): 4.0 - 6.0pH (1%, H2O): 4.0 - 6.0Note: This product is a general quality laboratory
chelating reagent suitable for most educational purposes. 0.1 M stock solutions of this grade may be pale yellow in color. For demanding research applications and for use with sensitive reagents, we recommend Ultrapure Grade (PN 15701) or ACS Reagent Grade EDTA (PN 15699).
CAS #6381-92-6
15697 1 kg
Ethylene Diamine Tetraacetic Acid, (EDTA) Tetrasodium Salt, DihydrateULTRAPURE
C10H12N2O8Na4•2H2O
Formula Weight: 416.20 (dihydrate)Product specifications:Form: White crystalline powderAssay: 99.0 - 101.0% (as is)Identity: By IRpH (1%, H2O): 10.7 - 11.7pH (5%, H2O): 10.7 - 12.0CAS #10378-23-1
15700 1 kg
Ethylene Glycol-O,O’-Bis (2 Amino-ethyl)-N,N,N’,N’,-Tetraacetic Acid (EGTA)ULTRAPURE
C14H24N2O10
Formula Weight: 380.35Product specifications:Form: White powderAssay: ≥ 99%Spectral Ratios (280 nm, 0.1 M in 1N NaOH, 1 cm):
≤ 0.10Identity: By IRLoss on Drying: ≤ 0.5%Residue on Ignition: ≤ 0.1%Chloride (Cl): ≤ 0.005%Sulfate (SO4): ≤ 0.02%Cadmium (Cd): ≤ 5 ppmCobalt (Co): ≤ 5 ppmCopper (Cu): ≤ 5 ppmIron (Fe): ≤ 5 ppmLead (Pb): ≤ 5 ppmNickel (Ni): ≤ 5 ppmZinc (Zn): ≤ 5 ppmApplication: � Chelating agent useful in the determination of
calcium in the presence of magnesiumReference:1. Schmid, R. W. and Reilley, C. N. (1957) Anal.
Chem. 29, 264-268.CAS #67-42-5
15703 10 gm 50 gm 100 gm
Fetal Bovine Serum (FBS)
Cell Culture GradeProduct specifications:Form: Clear straw to amber liquidTotal Protein: 3.0 - 4.0 gm/dlHemoglobin: ≤ 20 mg/dlpH: 6.7 - 8.0Endotoxin: ≤ 10 EU/mlSterility: SterileMycoplasma: None detectedVirally tested and found negative for BVD, IBR, and
PI3. USB Fetal Bovine Serum is an excellent supplement
to culture media.Storage: -20°C
15762 500 mI
Biochemicals
216 888-362-2447 | 216-765-5000 | usb.affymetrix.com
Flavin Adenine Dinucleotide, Disodium Salt, Hydrate(FAD)
C27H31N9O15P2Na2
Formula Weight: 829.51 (anhydrous) 883.56 (trihydrate)Product specifications:Form: Orange-yellow crystalline powderAssay (UV, 450 nm, pH 7.0): ≥ 95.0% (dry basis,
disodium salt)Moisture (KF): ≤ 8.0%Note: This product contains a varying amount of
moisture ranging from a dihydrate to a tetrahy-drate at the time of packaging.
Storage: 4°C, protect from light, dessicate.CAS #84366-81-4
15785 250 mg 500 mg 1 gm
Folic Acid(Pteroylglutamic Acid)
USBioAnalyzedC19H19N7O6
Formula Weight: 441.40Product specifications:Form: Yellow to yellow-orange crystalline powderAssay: 97.0 - 102.0% (dry basis)Chromatographic Purity: Passes testIdentity: Passes testMoisture (KF): ≤ 8.5%Residue on Ignition: ≤ 0.3%Organic Volatile Impurities: Meets requirementsStorage: Ambient, protect from light.CAS #59-30-3
15880 25 gm 100 gm 1 kg
FormamideULTRAPURE
Low ConductivityHCONH2
Formula Weight: 45.04Form: Clear, colorless liquidProduct specifications:Assay: ≥ 99%Refractive Index (20°C): 1.447 ± 0.001Density (20°C): 1.133 ± 0.001A (280 nm, 100%, 1 cm): ≤ 0.15A (260 nm, 100%, 1 cm): ≤ 2.3pH (50% v/v with H2O): 6.0 - 8.0Freezing Point: 2.0 - 3.0°CConductivity: ≤ 100 µmhos/cmCopper (Cu): ≤ 0.2 ppmIron (Fe): ≤ 1 ppmLead (Pb): ≤ 1 ppmZinc (Zn): ≤ 1 ppmStorage: 4°CApplications:� For use in nucleic acid hybridization buffers, spe-
cialized sequencing gels, and a variety of other molecular biology applications to reduce the Tm of nucleic acid hybrids
� Used to achieve stronger denaturing conditions in a sequencing gel along with 7-deaza-dGTP to resolve sequencing gel compressions.
To prepare a formamide sequencing gel: Place 40 ml Formamide in a beaker with mag-
netic stir bar. Acrylamide 7.6 gm N-N’-Methylene-bis-Acrylamide 0.4 gm Urea 42.0 gm 10X TBE Buffer 10 ml1. Cover with parafilm and warm in a 65°C water
bath with stirring until dissolved. This takes 30- 60 minutes and the mixture reaches a tempera-ture of about 50°C.
2. When completely dissolved, add water to a total volume of 100 ml.
3. Cover and continue stirring at room temperature until it reaches < 30°C.
4. Vacuum filter with a nitrocellulose filter.5. Add 1 ml of 10% ammonium persulfate and
0.15 ml of TEMED. Pour immediately. (Pouring this viscous mixture requires nearly a vertical angle for the glass plates.) The gel should be polymerized within 30 minutes.
Formamide gels should be run at the same power (wattage) as regular gels. This requires a higher voltage (60% higher) than normal gels, and DNA migrates about half as fast.
Soaking the gel in 5% acetic acid, 20% methanol helps prevent swelling. Soak 15 minutes for a 0.4 mm thick gel and dry normally.
Compression artifacts are caused whenever stable secondary structures exist in the DNA under the conditions prevailing in the gel matrix during electrophoresis.
The folded structure runs faster through the gel matrix than equivalent unfolded DNA, catch-ing up with the smaller DNA in the sequence. Currently, gel compression artifacts are elimi-nated in one of two ways. One is to change to a stronger denaturing condition, adding formamide as shown here, and the other is to use an analog of dGTP (dITP or 7-deaza-dGTP) during synthesis.
Formamide Hybridization Buffer: 50% Formamide, 5X SSC, 0.5% SDS, 5X
Denhardt’s Solution, 100 μg/ml sonicated fish sperm DNA
Reagent Amount for every 100 ml Formamide* 50 ml 20X SSC* 25 ml 20% SDS* 2.5 ml 50X Denhardt’s Solution* 10 ml 10 mg/ml fish DNA 1.0 ml Water* up to 100 ml*Denotes Ultrapure Reagent available.References:1. Sarkar, G., Kapelner, S. and Sommer, S. S. (1990)
Nucl. Acids Res. 18, 7465.2. Sambrook, J., Fritsch, E. F. and Maniatis, T. (1989)
Molecular Cloning, 2nd Edition, Cold Spring Harbor Laboratory.
CAS #75-12-7
75828 500 gm 1 kg
β-D-Fructose(β-D-Levulose)
C6H12O6
Formula Weight: 180.16Product specifications:Form: White crystalline powderAssay: 100 ± 2% (dry basis)Identity: By IRLoss on Drying: ≤ 0.5%Glucose: ≤ 0.5%Hydroxymethylfurfural: ≤ 0.1%Residue on Ignition: ≤ 0.5%Chloride (Cl): ≤ 0.018%Sulfate (SO4): ≤ 0.025%Arsenic (As): ≤ 1 ppmHeavy Metals (Pb): ≤ 5 ppmCAS #57-48-7
18425 1 kg
D-Fructose-6-Phosphate, Disodium Salt
C6H11Na2O9PFormula Weight: 304.10 (anhydrous) 322.11 (monohydrate)Practically free of Fructose-1,6-DiphosphateStorage: -20°CCAS #26177-86-6
15929 1 gm 5 gm
Biochemicals
217For bulk or alternate pack sizes, email us at [email protected].
G-418 Sulfate(Geneticin®, Antibiotic G-418)
ULTRAPURE
C20H40N4O10•2H2SO4
Formula Weight: 692.71Product specifications:Form: White powderPotency: ≥ 730 units/mg (dry basis)[α]25
D : +104° to 121° (c=1 in H2O)Moisture (KF): ≤ 6%Storage: 4°C
11379 500 mg 1 gm 5 gm
D-(+)-Galactose
C6H12O6
Formula Weight: 180.16Product specifications:Form: White to off-white powderAssay (HPLC): ≥ 99%Identity: By IR[α]20
D : ≥ +77° (c=4 in H2O and NH3)Loss on Drying: ≤ 0.5%Residue on Ignition: ≤ 0.5%CAS #59-23-4
15995 500 gm 1 kg 5 kg
GDP
See Guanosine-5’-Diphosphate, page 221
Gel Electrophoresis Reagents
Ethidium Bromide Drops, page 214Glycerol Tolerant Gel (GTG) Buffer, 20X Solution,
page 219RapidRun Agarose Buffer, 20X Solution, page 238TAE Buffer, 10X Solution, page 243TAE Buffer, 50X Solution, page 243TBE Buffer, 5X Solution, page 243TBE Buffer, 10X Ready-Mixed Powder, page 243Tris-Glycine (TG) Buffer, 10X Solution, page 247Tris-Glycine-SDS (TGS) Buffer, 10X Solution,
page 247
Gel Mixes and Solutions
Glycerol Tolerant Gel (GTG) Buffer, 20X Solution, page 219
RapidGel 40% Liquid Acrylamide Stock Solution, page 237
RapidGel-XL 6% DNA Sequencing Gel Solution, page 238
Stop Solution, page 242
Geneticin Disulfate
See G-418 Sulfate, page 217
Gentamicin Sulfate
USBioAnalyzedProduct specifications:Form: White to buff-colored powderActivity: ≥ 590 µg of gentamicin/mg (dry basis) ≥ 484 µg of gentamicin/mg (as is)Identity (A, B): Passes testspH (4%, H2O): 3.5 - 5.5[α]25
D : +107° to +121° (c=1 in H2O)Loss on Drying: ≤ 18.0%Residue on Ignition: ≤ 1.0%Gentamycin’s content: C1: 25 - 50% C2 + C2a: 25 - 55% C1a: 10 - 35%Limit of Methanol: ≤ 1.0%Mode of Action: Bactericidal, inhibits normal protein
synthesis.Applications:� Very stable� Autoclavable� Requires no refrigeration� Effective when combined with an antibiotic that
interferes with cell wall synthesis.CAS #1405-41-0
16051 1 gm 5 gm 25 gm
γ-Globulins, Fraction II
Source: Bovine BloodProduct specifications:Form: Lyophilized powderAssay: ≥ 98%Identity: By immuno-electrophoresisStorage: 4°CCAS #68476-36-8
16021 10 gm 25 gm
α-D-(+)-Glucose
See Dextrose, page 211
Glucose Oxidase
(E.C.1.1.3.4)Source: Aspergillus nigerForm: Stabilized, aqueous solutionActivity: Approx. 260 units/mg proteinProtein Concentration: Approx. 20 mg protein/mlContaminating Activities: Amylase: ≤ 0.05% Saccharase: ≤ 0.05% Maltase: ≤ 0.05%Unit Definition: One unit oxidizes 1 µmol of glucose/
minute at 25°C, pH 7.0.Storage: 4°C
16146 50,000 units 250,000 units 1 mu
D-Glucose-6-Phosphate, Disodium Salt, Dihydrate
C6H11Na2O9P•2H2OFormula Weight: 340.13 (dihydrate)Product specifications:Form: White powderAssay (enzymatic): ≥ 95%Storage: -20°C, desiccatedCAS #3671-99-6
16185 5 gm 10 gm
Glucose-6-Phosphate Dehydrogenase
(E.C.1.1.1.49)Source: Leuconostoc mesenteroidesProduct specifications:Form: Ammonium sulfate suspensionActivity: ≥ 400 units/mg protein (NAD units)Activity: ≥ 220 units/mg protein (NADP units)Protein Concentration: ≥ 8 mg/mlContaminating Activities: Hexokinase: ≤ 0.01% Phosphoglucose Isomerase: ≤ 0.005% Phosphogluconate Dehydrogenase: ≤ 0.001% Creatine Kinase: ≤ 0.001% Glutathione Reductase: ≤ 0.001% Phosphoglucomutase: ≤ 0.001% Myokinase: ≤ 0.001% Lactate Dehydrogenase: ≤ 0.01%Note: The above product is available in manufac-
turing quantities as a lyophilized powder or an ammonium sulfate suspension. Please inquire for bulk pricing.
Storage: 4°C
16171 1,000 units 5,000 units 10,000 units
Biochemicals
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Glucose-6-Phosphate Dehydrogenase
(E.C.1.1.1.49)Source: YeastProduct specifications:Form: Ammonium sulfate suspensionActivity: ≥ 250 units/mg proteinUnit Definition: One unit reduces 1 µmol NADP/
minute at 25°C, pH 7.8.Contaminating Activities: Glutathione Reductase: ≤ 0.2% Hexokinase: ≤ 0.02% Phosphogluconate Dehydrogenase: ≤ 0.01% Glucosephosphate Isomerase: ≤ 0.01% Phosphoglucomutase: ≤ 0.01% Creatine Kinase: ≤ 0.001% Myokinase: ≤ 0.01% ATPase: ≤ 0.001%Note: The above product is available in manufac-
turing quantities as a lyophilized powder or an ammonium sulfate suspension. Please inquire for bulk pricing.
Storage: 4°C
16176 1,000 units 5,000 units 10,000 units
Glucose-6-Phosphate Glutamate Dehydrogenase
(E.C.1.4.1.3)Source: Bovine LiverProduct specifications:Form: Lyophilized powderActivity: ≥ 10 units/mg powder, ~ 40 units/mg
enzyme proteinContaminating Activity: Malate dehydrogenase: < 0.01%Unit Definition: One unit catalyzes the oxidation of
1 µmol of 2-oxoglutarate/minute at 25°C under assay conditions not containing ADP.
Storage: -20°C
16250 100 mg 500 mg 1 gm
L-Glutamic Acid(L-α-Aminoglutaric Acid)
HOOCCH2CH2CH(NH2)COOHFormula Weight: 147.13Product specifications:Form: White crystalline powderAssay: 98.5 - 101.5% (dry basis)Identity: By IR[α]20
D : +31.2° to +32.2° (c=10 in 2N HCl)Loss on Drying: ≤ 0.1%Heavy Metals (Pb): ≤ 20 ppmArsenic (As): ≤ 1.5 ppmLead (Pb): ≤ 10 ppmCAS #56-86-0
16215 1 kg 5 kg 25 kg
L-Glutamic Acid, Monosodium Salt, Monohydrate(Monosodium Glutamate)
USBioAnalyzedC5H8NNaO4
•H2OFormula Weight: 187.13Product specifications:Form: White crystalline powderAssay: 99.0 - 100.5%Identity: By IRIdentification (A, B, C,): Passes testsClarity and color of solution: Passes testpH (5%, H2O): 6.7 - 7.2[α]20
D : +24.8° to +25.3° (c=10 in 2N HCl)Loss on Drying (100°C, 5 hours): ≤ 0.5%Heavy Metals (Pb): ≤ 20 ppmChloride (Cl): ≤ 0.25%Lead (Pb): ≤ 10 ppmResidual Solvents: Meets requirementsCAS #6106-04-3
16245 5 kg
L-Glutamine(L-α-Aminoglutamic Acid)
H2NCOCH2CH2CH(NH2)COOHFormula Weight: 146.14Product specifications:Form: White crystalline powderAssay: ≥ 98.5%Identity: By IR[α]20
D : +6.3° to +7.3° (c=4 in H2O)Loss on Drying: ≤ 0.2%Residue on Ignition: ≤ 0.1%Heavy Metals (Pb): ≤ 10 ppmArsenic (As): ≤ 3 ppmChloride (Cl): ≤ 0.021%Iron (Fe): ≤ 10 ppmSulfate (SO4): ≤ 0.028%CAS #56-85-9
16285 100 gm 250 gm 500 gm 1 kg
Glutamyl Cysteinyl Glycine
See Glutathione, page 218
Glutathione-Oxidized
C20H32O12N6S2•H2O
Formula Weight: 612.63Product specifications:Form: White lyophilized powderAssay (HPLC): ≥ 95%Identity: By IR[α]20
D : -92° to -106° (c=4 in H2O)Moisture (KF): ≤ 8%Heavy Metals (Pb): ≤ 10 ppmIron (Fe): ≤ 10 ppmStorage: 4°C, desiccatedCAS #27025-41-8
16320 250 mg 1 gm
Glutathione-Reduced(γ-L-Glutamyl-Cysteinyl Glycine)
C10H17O6N3SFormula Weight: 307.32Product specifications:Form: White crystalline powderAssay: ≥ 98%[α]20
D : -15° to -18° (c=2 in H2O)Loss on Drying: ≤ 0.5%Residue on Ignition: ≤ 0.2%Heavy Metals (Pb): ≤ 20 ppmArsenic (As): ≤ 2 ppmIron (Fe): ≤ 5 ppmCAS #70-18-8
16315 25 gm 100 gm
Biochemicals
219For bulk or alternate pack sizes, email us at [email protected].
Glycerin
See Glycerol, page 219
Glycerol(Glycerin)
ULTRAPURE
MB GradeHOCH2CH(OH)CH2OHFormula Weight: 92.09Product specifications:Form: Clear, colorless, viscous liquidAssay (LISP): 99.0 - 101.0%Identity: By IRSpecific Gravity (25°C): ≥1.249Color (APHA): ≤ 10Neutrality: Passes testResidue on Ignition: ≤ 0.007%Moisture: ≤ 0.5%Fatty Acid Esters: ≤ 0.3 ml of 0.5N NGOHSilver Reducing Substances: Passes testReadily Carbonizable Substances: Passes testRefractive Index: 1.470-1.475Chlorinated Compounds: ≤ 0.003%Chloride (Cl): ≤ 0.001%Sulfate (SO4): ≤ 0.002%Arsenic (As): ≤ 1.5 ppmCalcium (Ca): ≤ 5 ppmIron (Fe): ≤ 5 ppmMagnesium (Mg): ≤ 1 ppmMercury (Hg): ≤ 10 ppmZinc (Zn): ≤ 2 ppmHeavy Metals (Pb): ≤ 2 ppmDNase (endo): Passes testDNase (exo): Passes testRNase: Passes testProtease: Passes testApplications:� Used for long-term storage of bacterial cultures.� Step gradients for bacteriophage/particle purifica-
tion� Mammalian cell transfection� Storage buffers for enzymesTypical storage for bacterial cultures: Saturated bacterial culture 0.85 ml Glycerol, Ultrapure, sterile 0.15 ml Vortex the solution and transfer to a screw cap
storage tube. Freeze quickly and store at -70°C.CAS #56-81-5
16374 500 ml 1 L
Glycerol Tolerant Gel (GTG) Buffer, 20X SolutionULTRAPURE
MB GradeGlycerol Tolerant Gel (GTG) Buffer is formulated to resolve the problem of gel distortion associated with sequencing samples that contain glycerol. Standard TBE buffer causes gel distortion between 300- 500 bases as a result of reactions between glycerol and boric acid to form an anionic ester compound, which migrates in the sequencing gel. This buffer also works equally well with samples that contain no glycerol.Form: Solution consists of 1.78 M Tris, 0.57 M
Taurine, and 0.01 M EDTA. Solution is 0.2 µm fil-tered and dispensed into durable, square Nalgene bottles. Dilute to 1X with high purity dH2O for a solution consisting of 0.089 M Tris, 0.029 M Taurine, and 0.5 mM EDTA. Total volume is 1.5 liters of 20X buffer or 30 liters of 1X buffer.
pH (1:20, 25°C): 8.9 ± 0.1Conductivity 1:20, 830 ± 100 µmhos/cmTesting: Meets or exceeds functional test criteria for
high quality gel electrophoresis. Application: � Specifically formulated to alleviate the effects of
glycerol in DNA sequencing or PAGE gels.Storage: Ambient. Crystals which form in the cold at
low temperatures can be redissolved by warming briefly.
75827 1 L
β-Glycerophosphate, Sodium Salt, Pentahydrate
C3H5(OH)2PO4Na2•5H2O
Formula Weight: 306.11Alpha Content: 50%CAS #1555-56-2
21655 100 gm 500 gm 1 kg
GlycineULTRAPURE
MB GradeH2NCH2CO2HFormula Weight: 75.07Product specifications:Form: White powderAssay: ≥ 99.0%Identity: By IRλmax: 260 nm, 1 M, 1 cm: ≤ 0.05 280 nm, 1 M, 1 cm: ≤ 0.05Loss on Drying: ≤ 0.2%Residue on Ignition: ≤ 0.1%Readily Carbonizable Substances: Passes testHydrolyzable Substances: Passes testArsenic (As): ≤ 1 ppmBarium (Ba): ≤ 1 ppmCadmium (Cd): ≤ 1 ppmCalcium (Ca): ≤ 5 ppmChloride (Cl): ≤ 50 ppmCopper (Cu): ≤ 1 ppmIron (Fe): ≤ 1 ppmLead (Pb): ≤ 1 ppmMagnesium (Mg): ≤ 1 ppmManganese (Mn): ≤ 1 ppmSulfate (SO4): ≤ 50 ppmZinc (Zn): ≤ 1 ppmContaminant Testing: DNase (endo): Passes test DNase (exo): Passes test RNase: Passes testCAS #56-40-6
16407 500 gm 1 kg 5 kg
Turn to us as your single sourcingsolution for raw materials. Bulk quantities are available.
Biochemicals
220 888-362-2447 | 216-765-5000 | usb.affymetrix.com
Glycine
ACS Reagent GradeH2NCH2COOHFormula Weight: 75.07Product specifications:Form: White crystalline powderAssay: ≥ 98.5%Residue on Ignition: ≤ 0.1%Heavy Metals (Pb): ≤ 0.002%Chloride (Cl): ≤ 0.005%Sulfate (SO4): ≤ 0.005%Ammonium (NH4): ≤ 0.005%Substances Darkened by Sulfuric Acid: Passes testHydrolyzable Substances: Passes testCAS #56-40-6
16372 2.5 kg
Glycine
USBioAnalyzedH2NCH2CO2HFormula Weight: 75.07Product specifications:Form: White crystalline powderAssay: 98.5 - 101.5% (dry basis)Identity: By IRLoss on Drying: ≤ 0.2%Residue on Ignition: ≤ 0.1%Sulfate (SO4): ≤ 65 ppmHeavy Metals (Pb): ≤ 20 ppmChloride (Cl): ≤ 70 ppmHydrolyzable Substances: Passes testResidual Solvents: Meets requirementsCAS #56-40-6
16405 1 kg 5 kg 25 kg
GlycogenULTRAPURE
Source: OystersProduct specifications:Form: Light tan powderIdentity: By IR[α]20
D : +170° to 200° (c=0.2, H2O, dry basis)Solubility (0.25%, 1 M NaCl): Passes testLoss on Drying (110°C, 4 hours.): ≤ 15%CAS #9005-79-2
16445 5 gm 10 gm 25 gm
Glycogen, 20 mg/mlULTRAPURE
MB GradeSource: OystersProduct specifications:Concentration: 20 mg/ml in nuclease-free waterNote: Glycogen is an inert carrier which significantly
increases the efficiency of extracted DNA and RNA by ethanol precipitation.
Contaminant Testing: DNase (endo): Passes test DNase (exo): Passes test RNase: Passes testStorage: -20°C
77534 1 ml 5 x 1 ml
Glycyl-Glycine
NH2CH2CONHCH2COOHFormula Weight: 132.12Product specifications:Form: White crystalline powderAssay: ≥ 98.5%Chromatography (TLC): HomogeneousIdentity: By IRpKa (25°C): 8.25 ± 0.15Solubility (10%, H2O): Clear and colorlessLoss on Drying: ≤ 0.2%Chloride (Cl): ≤ 0.02%CAS #556-50-3
16505 100 gm 1 kg
GTP
See Guanosine-5’-Triphosphate, page 221
Guanidine HydrochlorideULTRAPURE
MB Grade(NH2)2C=NH•HClFormula Weight: 95.53Product specifications:Form: White/clear crystalsIdentity: By IRAssay: ≥ 99.5%Solubility (6 M, H2O): Clear and colorlessA (225 nm, 6 M, 1 cm): ≤ 0.2A (260 nm, 6 M, 1 cm): ≤ 0.03Melting Point: 184 ± 3°CResidue on Ignition: ≤ 0.05%Aldehyde: None detectedAmmeline: None detectedArsenic (As): ≤ 1 ppmCopper (Cu): ≤ 1 ppmIron (Fe): ≤ 2 ppmLead (Pb): ≤ 5 ppmSulfate (SO4): ≤ 50 ppmZinc (Zn): ≤ 1 ppmDNase (endo): Passes testRNase: Passes testProtease: Passes testApplications:� Used for extraction of RNA.� Preparation of high molecular weight genomic
DNA� Solubilizing inclusion bodiesPhenol/Chloroform RNA Isolation MethodA. Guanidine HCl Homogenization Buffer for RNA
Isolation: 8.0 M Guanidine HCl, 0.1 M Sodium Acetate, pH 5.5, 5.0 mM Dithiothreitol, 0.5% Sodium Lauryl Sarcosinate
B. Extraction with Phenol: Most extractions are per-formed using equal volumes of equilibrated phe-nol with respect to the sample volume. Normally, the extraction is performed twice with phenol and once with a 1:1 solution of phenol/chloro-form. Hence, it is estimated that 25 ml of phenol is required for every 10 ml of sample.
Note: If regular phenol is purchased, it will need to be equilibrated before use. For this reason, many customers may prefer to purchase phenol that is already equilibrated (PN 75829).
C. RNA Resuspension Buffer: TE, pH 7.5.CAS #50-01-1
75823 100 gm 500 gm
Biochemicals
221For bulk or alternate pack sizes, email us at [email protected].
Guanidine Hydrochloride
(NH2)2C=NH•HClFormula Weight: 95.53Product specifications:Form: White/clear crystalsIdentity: By IRMelting Point: 184 - 190°CLoss on Drying: ≤ 0.50%Note: Contains some water-insoluble matter.CAS #50-01-1
34007 1 kg 5 kg
Guanidine ThiocyanateULTRAPURE
(NH2)2C=NH•HNCSFormula Weight: 118.16Product specifications:Form: White crystalline powderAssay: ≥ 99%Identity: By IRA (280 nm, 6 M, 1 cm): ≤ 0.8Solubility (5%, H2O): Passes testSolubility (5%, MeOH): Passes testMelting Point: 120 ± 2°CMoisture (KF): ≤ 1%Copper (Cu): ≤ 0.5 ppmIron (Fe): ≤ 5 ppmLead (Pb): ≤ 1 ppmZinc (Zn): ≤ 2 ppmApplications:� Used to extract RNA from cells and tissue.� Denatures protein, thus inactivating ribonu-
cleases.Solution for Guanidine Thiocyanate RNA Extraction: Guanidine Thiocyanate 4.0 M Dithiothreitol 0.1 mM Sodium Acetate, pH 5.2 20 mM N-Lauryl sarcosine 0.5% Heat slightly (65°C) to dissolve the guanidine
thiocyanate and sodium acetate. Add the dithio-threitol and N-lauryl sarcosine. The pH should be about 5.5. Filter and store at room temperature.
Cesium Chloride RNA Isolation MethodA. Guanidine Thiocyanate Homogenization Buffer
for RNA Isolation: 4.0 M Guanidine Thiocyanate, 0.1 M Tris, pH 7.5, 1.0% 2-mercaptoethanol.
B. Cesium Chloride RNA Isolation Solution: Most researchers layer their RNA homogenates onto a cushion of 5.7 M Cesium Chloride.
C. RNA Resuspension Buffer: TE, pH 7.5.CAS #593-84-0
75818 100 gm 250 gm 1 kg
Guanosine-5’-Diphosphate, Disodium Salt(GDP)
C10H13N5O11P2Na2•3H2O
Formula Weight: 541.21Reference constants:λmax (pH 7, NRC): 253 nmεmax (pH 7, NRC): 13.7 x 103
ε260 (pH 7, DBR): 11.8 x 103
Storage: -20°C, protect from moisture.CAS #43139-22-6
16785 100 mg 500 mg 1 gm
Guanosine-5’-Triphosphate, Sodium Salt, 100 mM Solution(GTP)
ULTRAPURE
C10H12N5O14P3Na4
Formula Weight: 611.11Form: 100 mM aqueous solution, pH 7.5Storage: -20°C
77243 25 µmol (250 µl)
Guanosine-5’-Triphosphate, Disodium Salt(GTP)
C10H14N5O14P3Na2
Formula Weight: 567.14Product specifications:Assay: ≥ 95%λmax: 252 nm ± 1nmεmax: (13.7 ± 0.7) x103
Spectral Ratios (pH 7): A250/A260: 1.15 ± 0.03 A280/A260: 0.65 ± 0.03Moisture (KF): ≤ 10.0%Reference constants:λmax (pH 7, NRC): 252 nmεmax (pH 7, NRC): 13.7 x 103
ε260 (pH 7, DBR): 11.7 x 103
Storage: -20°C in tightly closed containers.CAS #56001-37-7
16800 25 mg 100 mg 500 mg 1 gm
Hemin
Source: PorcineC34H32O4N4FeCl Formula Weight: 651.94 Product specifications:Form: Dark blue/purple to black crystalline powderAssay: ≥ 96%Identity: By IRMoisture (KF): ≤ 2.0%Solubility (0.02%, 0.1N NaoH): Passes testCAS #16009-13-5
16880 10 gm 50 gm
Heparin, Sodium Salt
USBioAnalyzedProduct specifications:Form: White to pale cream colored powder.Assay: ≥ 180 USP Heparin units/mg (dry basis)Identification A: Passes testIdentification B: Passes testIdentification C: 0.9 - 1.1Identification D: Passes testAnti-Factor Xa Activity: Record USP U/mgAnti-Factor IIa Potency (as is): Record USP U/mgResidue on Ignition: 28.0 to 41.0%Nitrogen Content: 1.3 to 2.5% (dry basis)Heavy Metals: ≤ 0.003% Organic Impurities Limit of Galactosamine: ≤ 1%Nucleotidic Impurities: ≤ 0.20Absence of Oversulfated Chondroitin Sulfate A:
Passes testAbsence of Oversulfated Chondroitin Sulfate B:
Passes testProtein Impurities: ≤ 1.0%pH (1%, H2O): 5.0 to 7.5Bacterial Endotoxins: ≤ 0.03 Endotoxin units/
Heparin unitLoss on Drying: ≤ 5.0% Residual Solvents: Passes TestStorage: Ambient, in tightly closed containers.CAS #9041-08-1
16920 50,000 units 100,000 units 500,000 units 1 mu
Biochemicals
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HEPES, 1 M Solution, pH 7.3ULTRAPURE
MB GradeForm: 1 M solution of HEPES in high purity H2O, pH
7.3 (at 37°C). Solution is 0.2 µm filtered and dis-pensed into durable, square Nalgene bottles.
pH (37°C): 7.3 ± 0.1Contaminant Testing: DNase (endo): Passes test DNase (exo): Passes test RNase: Passes test Protease: Passes testStorage: Ambient, store at +4°C after opening.
16924 100 ml 500 ml 1 L
HEPES(N-[2-Hydroxyethyl]piperazine-N’ [2-Ethane-sulfonic Acid])
ULTRAPURE
C8H18N2O4SFormula Weight: 238.30Product specifications:Form: White crystalline powderAssay: ≥ 99.0% (dry basis)Identity: By IRpKa (20°C): 7.55 ± 0.15A (250 nm, 1 M, 1 cm): ≤ 0.2Loss on Drying: ≤ 0.5%Residue on Ignition: ≤ 0.1%Chloride (Cl): ≤ 0.005%Phosphate (PO4): ≤ 0.001%Sulfate (SO4): ≤ 0.01%Iron (Fe): ≤ 5 ppmHeavy Metals (Pb): ≤ 10 ppmCAS #7365-45-9
16926 100 gm 250 gm 1 kg 5 kg
HEPES, Sodium Salt
HO(CH2)2(C4H8N2)(CH2)2SO3NaFormula Weight: 260.29Product specifications:Form: White crystalline powderAssay: ≥ 98.0% (dry basis)Identity: By IRpKa (20°C): 7.55 ± 0.15pH (0.1 M, H2O): 10.2 ± 0.8A (250 nm, 5%, 1 cm): ≤ 0.30A (260 nm, 5%, 1 cm): ≤ 0.175Solubility (5%, H2O): Clear solutionLoss on Drying: ≤ 4.0%Heavy Metals (Pb): ≤ 10 ppmStorage: Hygroscopic, store in tightly closed con-
tainer.CAS #75277-39-3
16928 100 gm 1 kg
Hexokinase
(E.C.2.7.1.1)Source: YeastProduct specifications:Form: Lyophilized, ammonium sulfate-free powderActivity: ≥ 140 units/mg protein ≥ 80 units/mg powderUnit Definition: One unit converts 1 µmol of glucose
to glucose-6-phosphate per minutes at 25°C, pH 7.5 in the presence of ATP.
Contaminating Activities: PGI: ≤ 0.1% G-6-PDH: ≤ 0.01% PGluM: ≤ 0.01% MK: ≤ 0.01% ATPase: ≤ 0.003% PGDH: ≤ 0.005% CK: ≤ 0.005% GR: ≤ 0.05%Note: The above product is available in manufac-
turing quantities as a lyophilized powder or an ammonium sulfate suspension. Please inquire for bulk pricing.
Storage: -20°C, protect from moisture.CAS #9001-51-8
16965 1,000 units 10,000 units 25,000 units 50,000 units
L-Histidine, Free Base
USBioAnalyzed(C3N2H3)CH2CH(NH2)COOHFormula Weight: 155.15Product specifications:Form: White crystalline powderAssay: 98.5 - 101.5% C6H9N3O2 (dry basis)Identity: By IRChromatographic Purity: Passes testpH (2%, H2O): 7.0 - 8.5[α]25
D : +12.6° to +14.0° (c=11 in 6N HCl)Loss on Drying: ≤ 0.2%Residue on Ignition: ≤ 0.4%Heavy Metals (Pb): ≤ 0.0015%Chloride (Cl): ≤ 0.05%Iron (Fe): ≤ 0.003%Sulfate (SO4): ≤ 0.03%CAS #71-00-1
17070 100 gm 1 kg
L-Histidine, Monohydrochloride, Monohydrate
(C3H3N2)CH2CH(NH2)COOH•HCl•H2OFormula Weight: 209.63Product specifications:Form: White crystalline powderAssay: ≥ 98.0%Identity: By IRChromatography (TLC): Homogeneous[α]20
D : +9.2° to +10.6° (c=11 in 6N HCl)Loss on Drying: ≤ 0.3%Residue on Ignition: ≤ 0.10%Heavy Metals (Pb): ≤ 10 ppmAmmonium (NH4): ≤ 0.02%Arsenic (As2O3): ≤ 1 ppmIron (Fe): ≤ 30 ppmSulfate (SO4): ≤ 0.03%CAS #5934-29-2
17080 100 gm 1 kg 5 kg
L-Hydroxysuccinic Acid
See L-Malic Acid, page 227
Biochemicals
223For bulk or alternate pack sizes, email us at [email protected].
ImidazoleULTRAPURE
C3H4N2
Formula Weight: 68.08Product specifications:Form: White to cream crystalline flakes/powderAssay: ≥ 98.5% (dry basis)Identity: By IRpKa (25°C): 6.95 ± 0.15Melting Point: 88 - 92°CMoisture: ≤ 1.0%Iron (Fe): ≤ 30 ppmHeavy Metals (Pb): ≤ 10 ppmCAS #288-32-4
17525 100 gm 500 gm 1 kg 5 kg
IPTG
See Isopropyl-β-D-Thiogalactopyranoside, page 223
Isocitrate Dehydrogenase, (NADP+ Specific), Recombinant[r-ICDH (NADP+)]
NADP+ Specific(E.C.1.1.1.42)Source: Yeast recombinant in S. cerevisiaeProduct specifications:Form: 50% Glycerol SolutionSpecific Activity: ≥ 30 units/mg proteinpH: 7.0 ± 0.5Unit Definition: One unit converts 1 µmol of isoci-
trate to α-Ketoglutarate/minute at 25°C, pH 8.5.Contaminating Activity: ICDH (NAD+): ≤ 0.5%Note: The above product is available in manufactur-
ing quantities as an ammonium sulfate suspen-sion or a glycerol solution. Please inquire for bulk pricing.
Storage: -20°C
17798 150 units 600 units
DL-Isoleucine
CH3CH2CH(CH3)CH(NH2)COOHFormula Weight: 131.17Product specifications:Form: White crystalline powderAssay: ≥ 98.0%Identity: By IRChromatography (TLC): HomogeneousLoss on Drying: ≤ 0.3%Residue on Ignition: ≤ 0.1%Heavy Metals: ≤ 20 ppmChloride (Cl): ≤ 0.02%Iron (Fe): ≤ 50 ppmSulfate (SO4): ≤ 0.05%CAS #443-79-8
17820 100 gm 500 gm 1 kg
L-Isoleucine
USBioAnalyzedCH3CH2CH(CH3)CH(NH2)COOHFormula Weight: 131.17Product specifications:Form: White crystalline powderAssay: 98.5 - 101.5% C6H13NO2 (dry basis)Identity: By IRChromatographic Purity: Passes testpH (1%, H2O): 5.5 - 7.0[α]20
D : +38.9° to +41.8° (c=4 in 6N HCl, 20°C)Loss on Drying: ≤ 0.3%Residue on Ignition: ≤ 0.3%Sulfate (SO4): ≤ 0.03%Heavy Metals (Pb): ≤ 0.0015%Chloride (Cl): ≤ 0.05%Iron (Fe): ≤ 0.003%CAS #73-32-5
17825 10 gm 100 gm 500 gm 1 kg
Isopropyl-β-D-Thiogalactopyranoside (IPTG), Dioxane FreeULTRAPURE
C9H18O5SFormula Weight: 238.30Product specifications:Form: White to off-white powderAssay: ≥ 99%Identity: By IR[α]20
D : -31.5°C ± 1° (c=1 in H2O)Melting Point: 109 - 114°CMoisture (KF): ≤ 2.0%Dioxane (GC): ≤ 0.02%Applications:� Induces the lac operon in E. coli by binding and
inactivating the lac repressor.� Selection for lac Y mutants� Selection for recombinant plasmids containing
β-galactosidase fusion proteins� Induces the cellular content of lactose permease.Storage: -20°C, desiccated, protect from light.CAS #367-93-1
17886 1 gm 5 gm 25 gm 100 gm
Kanamycin Sulfate
C18H36N4O11•H2SO4
Formula Weight: 582.58Product specifications:Form: White to off-white powderPotency: ≥ 750 µg of C18H36N4O11/mg powder (dry basis)Identity (A, B, C): Passes testspH (1%, H2O): 6.5 - 8.5Loss on Drying: ≤ 1.5%Residue on Ignition: ≤ 0.5%Specific Rotation: +112° ± 123° (c=1 in H2O)Mode of Action: Binds to ribosomal components
and inhibits protein synthesis. Kanamycin is inac-tivated by an aminoglycoside phospho transfer-ase (MW 25,000) in the periplasmic space.
Application:� A selectable marker for cosmid vectors used in
transfection of mammalian cells.Storage: 4°CCAS #25389-94-0
17924 5 gm 25 gm
Biochemicals
224 888-362-2447 | 216-765-5000 | usb.affymetrix.com
α-Ketoglutaric Acid, Free Acid
C5H6O5
Formula Weight: 146.10Product specifications:Form: White to off-white crystalline powderAssay: ≥ 98.0% (as is)Identity: By IRMelting Point: 114 - 119°CLoss on Drying: ≤ 1.0%CAS #328-50-7
17950 100 gm 500 gm 1 kg
α-Ketoglutaric Acid, Disodium Salt, Dihydrate
C5H4O5Na2•2H2O
Formula Weight: 226.09 (dihydrate) 190.06 (anhydrous)Product specifications:Form: White powderAssay: ≥ 98.0% (as is)Identity: By IRMoisture: 14.0 - 18.0%CAS #305-72-6
17955 5 gm 100 gm 1 kg
L- (+)-Lactic Acid, Lithium Salt(Lithium Lactate)
C3H5LiO3
Formula Weight: 96.01Product specifications:Form: White crystalline powder[α]20
D : -11.5° ± 1.5° (c=10, H2O)Solubility: Passes testLoss on Drying: ≤ 0.5%CAS #27848-80-2
18160 100 gm
Lactoferrin
Form: Off-white powderPurity: ≥ 95%Protein (as is): ≥ 93%Ash: ≤ 1.5%Storage: 4°C, in tightly closed containers.
18177 100 mg 500 mg 1 gm
D-(+)-Lactose, Monohydrate
USBioAnalyzedC12H22O11
•H2OFormula Weight: 360.31Product specifications:Form: White powderClarity and Color of Solution: Passes testIdentity (A, B, C): Passes tests[α]20
D : +54.4° to +55.9° (c=10 in 0.2 ml 6N NH4OH/100 ml H2O)
Acidity or Alkalinity: Passes testLoss on Drying: ≤ 0.5%Moisture (KF): 4.5 - 5.5%Residue on Ignition: ≤ 0.1%Protein and Light-Absorbing Impurities: Passes testHeavy Metals (Pb): ≤ 5 ppmMicrobial Limits: Passes testCAS #64044-51-5
18185 5 kg
N-Lauroyl Sarcosine, Sodium Salt(Sodium-N-Lauryl Sarcosine)
ULTRAPURE
HO2CCH2CH3NCO(CH2)10CH2•Na
Formula Weight: 293.38Product specifications:Form: White to faint white-yellow powderIdentity: By IRMoisture (KF): ≤ 5.0%pH (10%, H2O): 7.5 - 8.5Solubility (10%, H2O): Clear to slightly hazy, color-
less to faint yellow solutionCAS #137-16-6
21653 50 gm 100 gm
Lauryl Sulfate, Lithium Salt
See Lithium Dodecyl Sulfate, page 225
Lauryl Sulfate, Sodium Salt
See Sodium Dodecyl Sulfate, page 240
LB Agar, Ready-Made Powder
Form: Off-white free flowing powder.pH (3.5%, H2O): 7.0 ± 0.2Contents: Casein Peptone: 10 gm/L Yeast Extract: 5 gm/L Sodium Chloride: 5 gm/L Agar: 15 gm/LTo use: Dissolve 35 gm powder per liter of purified
water. Autoclave, cool and make plates.Application: � For the propagation of bacteria
75851 250 gm 1 kg
LB Broth, Ready-Made Powder
Form: Luria-Bertani Broth. Off-white free flowing powder.
pH (2.0%, H2O): 7.0 ± 0.2Contents: Casein Peptone: 50% Yeast Extract: 25% Sodium Chloride: 25%To use: Dissolve 20 gm powder per liter of purified
water. Autoclave.Application: � For the propagation of bacteria
75852 250 gm 1 kg
Lecithin
Source: SoybeanProduct specifications:Form: Off-white to light tan granular powderMoisture: ≤ 1.5% (at time of packaging)Acetone Insolubles: ≥ 95.0%Hexane Insolubles: ≤ 0.3%CAS #8002-43-5
18245 500 gm 1 kg
Leptomycin B
C33H48O6
Formula Weight: 540.73Form: Leptomycin B is an antifungal antibiotic that
blocks CRM1 mediated nuclear export of proteins containing Nuclear Export Signal (NES). It inac-tivates CRM1 by covalently modifying the sulf-hydryl residue of Cysteine-529. This inhibits the formation of a complex between CRM1, Ran, and a protein with a NES. This results in the nuclear retention of the NES-bearing protein(1). Inhibiting the CRM1-mediated nucleo-cytoplasmic export of proteins bearing a NES produces the nuclear accumulation of a large number of proteins(2) such as Actin, c-Abl, Cdc25, Cyclin B1, HDM2, HIV-1, IΚBα, and p53.
References1. Kudo, N., Matsumori, N., Taoka, H., Fujiwara,
D., Schreiner, E. P., Wolff, B., Yoshida, M. and Horinouchi, S. (1999) Proc. Natl. Acad. Sci. USA 96, 9112-9117.
2. Lain, S., Xirodimas, D. and Lane, D. (1999) Experimental Cell Research 253, 315-324.
CAS #87081-35-4
25110 1 µg
Biochemicals
225For bulk or alternate pack sizes, email us at [email protected].
DL-Leucine
(CH3)2CHCH2CH(NH2)COOHFormula Weight: 131.17Product specifications:Form: White crystalline powderAssay: ≥ 98.0% (as is)Identity: By IRChromatography (TLC): HomogeneousLoss on Drying: ≤ 0.3%CAS #328-39-2
18270 100 gm 1 kg
L-Leucine
C6H13NO2
Formula Weight: 131.17Product specifications:Form: White crystalline powderAssay: 98.5 - 101.5% (dry basis)Identity: By IR[α]20
D : +14.5° to +16.5° (c=4 in 6N HCl dry basis)Loss on Drying: ≤ 0.2%Residue on Ignition: ≤ 0.1%Lead (Pb): ≤ 10 ppmHeavy Metals (Pb): ≤ 20 ppmCAS #61-90-5
18280 100 gm 1 kg 5 kg
LeupeptinULTRAPURE
C20H38N6O4•1⁄2 H2SO4
Formula Weight: 475.59Form: White powderApplications:� Leupeptin, a protease inhibitor, will strongly
inhibit Trypsin, Papain, Plasmin, Thrombokinase, Kallikrein, and Cathepsin B. The half-maximal inhibitory concentration ranges from 0.5 to 75 µg/ml, depending on the enzyme and the substrate.
� Leupeptin does not inhibit Chymotrypsin, Elastase, Renin, or Pepsin.
Storage: -20°CCAS #55123-66-5
18413 5 mg 25 mg 100 mg
DL-α-Lipoic Acid(DL-6,8-Thioctic Acid)
C8H14O2S2
Formula Weight: 206.33Product specifications:Form: Light yellow crystalline powderAssay: ≥ 98.0% (dry basis)Identity: Passes testLoss on Drying: ≤ 0.5%Storage: In tightly closed, light-resistant containers.CAS #1077-28-7
18505 25 gm 50 gm 100 gm
DL-α-Lipoic Acid (DL-6,8-Thioctic Acid)
C8H14O2S2
Formula Weight: 206.33Product specifications:Form: Light yellow crystalline powderAssay: ≥ 98.0% (dry basis)Identity: By IRMoisture (KF): ≤ 0.5%Melting Point: 60-62°CSolubility ( 3 gm/60 ml EtOH): Light yellow solutionStorage: Ambient, in tight, light resistant containersCAS# 1077-28-7
18522 50 gm 100 gm 1 kg
Lithium Chloride (LiCl), 7.5 M SolutionULTRAPURE
MB GradeForm: 7.5 M solution of lithium chloride (LiCl) in
high purity dH2O. Solution is 0.2 µm filtered and dispensed into durable, square Nalgene bottles.
Contaminant Testing: DNase (endo): Passes test DNase (exo): Passes test RNase: Passes testCAS #7447-41-8
75902 100 ml
Lithium Dodecyl Sulfate(Lithium Lauryl Sulfate)
ULTRAPURE
CH3(CH2)11OSO3LiFormula Weight: 272.33Product specifications:Assay (acidity): ≥ 98%Assay (C12 by GC): ≥ 98%A (230 nm, 3%, 1 cm): ≤ 0.2A (280 nm, 3%, 1 cm): ≤ 0.1Loss on Drying: ≤ 1%Chloride (Cl): ≤ 0.01%Copper (Cu): ≤ 5 ppmIron (Fe): ≤ 1 ppmLead (Pb): ≤ 5 ppmCAS #2044-56-6
32816 25 gm 100 gm
Lithium Lactate
See L-(+)-Lactic Acid, Lithium Salt, page 224
Lithium Lauryl Sulfate
See Lithium Dodecyl Sulfate, page 225
Luria Agar, Ready-Made Powder
Form: Off-white free flowing powderpH (4.0%, H2O): 7.0 ± 0.2Contents: Casein Peptone: 25% Yeast Extract: 12.5% Sodium Chloride: 2.5% Agar: 37.5%To use: Dissolve 40 gm powder per liter of purified
water. Autoclave, cool and make plates.Application: � For the propagation of bacteria
75853 250 gm 1 kg
Luria Broth, Ready-Made Powder
Form: Off-white free flowing powderpH (2.5%, H2O): 7.0 ± 0.2Contents: Casein Peptone: 40% Yeast Extract: 20% Sodium Chloride: 40%To use: Dissolve 25 gm powder per liter of purified
water. Autoclave.Application: � For the propagation of bacteria
75854 250 gm 1 kg
Biochemicals
226 888-362-2447 | 216-765-5000 | usb.affymetrix.com
L-Lysine, Hydrochloride
NH2(CH2)4CH(NH2)COOH•HClFormula Weight: 182.65Product specifications:Form: White crystalline powderAssay: ≥ 98.5% (dry basis)Identity: By IR[α]20
D : +19.0 to +21.5° (c=8 in 6N HCl)Loss on Drying: ≤ 0.5%Residue on Ignition: ≤ 0.1%Heavy Metals (Pb): < 10 ppmAmmonium (NH4): ≤ 0.02%Arsenic (As2O3): ≤ 1 ppmIron (Fe): ≤ 20 ppmSulfate (SO4): ≤ 0.03%CAS #657-27-2
18585 1 kg 5 kg 10 kg
LysozymeULTRAPURE
(E.C.3.2.1.17)Source: Chicken Egg WhiteProduct specifications:Form: Lyophilized powderActivity: Approximately 20,000 units/mgUnit Definition: One unit causes a decrease in
absorbance of 0.001 per minute at 450 nm, 25°C and pH 7.0 with Micrococcus lysodeikticus as a substrate.
Moisture: ≤ 6.0%Chloride: 3.2 - 4.2%pH (2%, H2O): 3.0 - 4.0Storage: -20°C, desiccated
18645 5 gm 10 gm 25 gm 100 gm
M13mp18, Single-Stranded DNA
See Molecular Biology Products & Kits section, page 129
Magnesium Chloride (MgCl2), 1 M SolutionULTRAPURE
MB GradeForm: 1 M solution of magnesium chloride (MgCl2)
in high purity dH2O. Solution is 0.2 µm filtered and dispensed into vials and/or durable, square Nalgene bottles.
Assay: 1.00 M ± 0.01 M (EDTA titration)Contaminant Testing: Heavy Metals(Pb): ≤ 5 ppm DNase (endo): Passes test DNase (exo): Passes test RNase: Passes test Protease: Passes test
78641 10 x 1 ml 100 ml
Magnesium Chloride, HexahydrateULTRAPURE
ACS Reagent GradeMgCl2•6H2OFormula Weight: 203.30Product specifications:Form: White crystalline powderAssay: 99.0 - 102.0%Insoluble Matter: ≤ 0.005%Nitrate (NO3): ≤ 0.001%Phosphate (PO4): ≤ 5 ppmSulfate (SO4): ≤ 0.002%Ammonium (NH4): ≤ 0.002%Barium (Ba): ≤ 0.005%Calcium: ≤ 0.01%Manganese (Mn): ≤ 5 ppmPotassium (K): ≤ 0.005%Sodium (Na): ≤ 0.005%Strontium (Sr): ≤ 0.005%Heavy Metals (Pb): ≤ 5 ppmIron (Fe): ≤ 5 ppmCAS #7791-18-6
18641 500 gm 1 kg
Malate Dehydrogenase, Lyophilized
(E.C.1.1.1.37)Source: Porcine Heart (Mitochondrial)Product specifications:Form: Lyophilized powder, ammonium sulfate freeActivity: ≥ 1,100 units/mg protein.Protein: 10 - 50%Unit Definition: One unit causes the oxidation of 1 µmol of NADH/minute at 25°C, pH 7.4.Contaminating Activities: Fumarase (L-Malate): ≤ 0.01% LDH: ≤ 0.01% GOT: ≤ 0.01% GDH (NAD): ≤ 0.002% NADH Oxidase: ≤ 0.001%Note: The above product is available in manufactur-
ing quantities as a lyophilized powder, an ammo-nium sulfate suspension or a glycerol solution. Please inquire for bulk pricing.
Storage: -20°CCAS #9001-64-3
18670 25,000 units 100,000 units
Maleic Acid(Toxilic Acid)
Reagent GradeC4H4O4
Formula Weight: 116.07Product specifications:Form: White crystalline powderAssay: 99.0 - 101.0% (dry basis)Identity: By IRSolubility (50%, H2O): Clear and colorlessMoisture (KF): ≤ 2.0%Iron (Fe): ≤ 5 ppmHeavy Metals (Pb): ≤ 10 ppmCAS #110-16-7
18685 1 kg
DL-Malic Acid
HOOCCH(OH)CH2COOHFormula Weight: 134.09Product specifications:Form: White granular powderAssay: ≥ 98% (as is)Identity: By IRMoisture (KF): ≤ 1.0%CAS #617-48-1
18695 1 kg
Biochemicals
227For bulk or alternate pack sizes, email us at [email protected].
L-Malic Acid
C4H6O5
Formula Weight: 134.09Product specifications:Form: White crystalline powderAssay (GC): ≥ 99%Identity: By IR[α]20
D : -26.5° to -30.0° (c=5.5 in pyridine)Melting Point: 102 - 106°CMoisture (KF): ≤ 1.0%CAS #97-67-6
18700 500 gm 1 kg
Maltose, Monohydrate
High Purity GradeC12H22O11
•H2OFormula Weight: 360.31Product specifications:Form: White crystalline powderAssay (HPLC): ≥ 98%Identity: By IR[α]20
D : +130.4° ± 1.3° (c=4 in H2O, NH4OH)Solubility (10%, H2O): Passes testLoss on Drying: ≤ 6%Residue on Ignition: ≤ 0.05%Dextrins: Passes testArsenic (As): ≤ 0.5 ppmIron (Fe): ≤ 5 ppmHeavy Metals (Cu): ≤ 10 ppmCAS #6363-53-7
18724 500 gm 1 kg
D-(+)-Maltose, Monohydrate
C12H22O11•H2O
Formula Weight: 360.31Product specifications:Form: White to off-white powderMaltose Content: ≥ 92.0% (dry basis)Glucose Content: ≤ 3.0% (dry basis)Loss on Drying: ≤ 7.0%Heavy Metals (Pb): ≤ 5 ppmCAS #6363-53-7
18725 1 kg 5 kg
Mannitol
USBioAnalyzedC6H14O6
Formula Weight: 182.17Product specifications:Form: White crystalline powderAssay: 96.0 - 101.5% (dry basis)Identity: Passes test[α]25
D : +137° to +145° (c=1 in USP prep.)Melting Point: 164 - 169°CLoss on Drying: ≤ 0.3%Acidity: Passes testReducing Sugars: Passes testChloride (Cl): ≤ 0.007%Sulfate (SO4): ≤ 0.01%Arsenic (As): ≤ 1 ppmCAS #69-65-8
18755 500 gm 1 kg 5 kg
D-(+)-Mannose
C6H12O6
Formula Weight: 180.16Product specifications:Form: White crystalline powderIdentity: By IR[α]20
D : +14.2° ± 0.7° (c=4 in H2O, NH3)Loss on Drying: ≤ 0.5%CAS #3458-28-4
18775 100 gm 500 gm
MES, Anhydrous(2-(N-Morpholino) Ethane Sulfonic Acid)
C6H13NO4SFormula Weight: 195.24CAS #4432-31-9
18888 100 gm 250 gm 1 kg
MES, Monohydrate(2-(N-Morpholino) Ethane Sulfonic Acid)
ULTRAPURE
HO3S(CH2)2(C4H8NO)•H2OFormula Weight: 213.25Product specifications:Form: White/clear crystalline powderAssay: ≥ 98.5% (dry basis)Identity: By IRpKa (20°C): 6.15 ± 0.15A (250 nm, 1 M, 1 cm): ≤ 0.1Moisture (KF): ≤ 11%Iron (Fe): ≤ 30 ppmLead (Pb): ≤ 10 ppmSolubility (1 M, H2O, 100 ml): Clear and colorlessNote: This product varies up to a monohydrate.CAS #145224-94-8
18886 100 gm 1 kg
MES, Sodium Salt(2-(N-Morpholino) Ethane Sulfonic Acid, Sodium Salt)
C6H12NO4SNaFormula Weight: 217.22Product specifications:Form: White/clear crystalline powderAssay: ≥ 98.5% (dry basis)Identity: By IRpKa (20°C): 6.15 ± 0.15A (260 nm, 1 M, 1 cm): ≤ 0.1Moisture (KF): ≤ 1%Iron (Fe): ≤ 30 ppmLead (Pb): ≤ 10 ppmSolubility (1 M, H2O, 100 ml): Clear and colorlessCAS #71119-23-8
18893 100 gm 250 gm 1 kg
Visit usb.affymetrix.com for new products, protocols, and MSDS documents.
Biochemicals
228 888-362-2447 | 216-765-5000 | usb.affymetrix.com
Methylene Blue, Trihydrate(Methylthionine Chloride)
(C.I.52015)C16H18ClN3S•3H2OFormula Weight: 373.90Product specifications:Form: Dark green crystalline powderAssay: 98.0 - 103.0% (dry basis)Identity: By IRλmax: 664 nm ± 1Solubility (1%, H2O): Passes testpH (1%, H2O): 3.6 - 4.5Moisture (KF): 8.0 - 18.0%Application:� Used in staining RNA that is bound to nylon
and nitrocellulose membranes. Methylene blue is the stain of choice in visualizing filter bound RNA over a pre-transfer gel stain with ethidium bromide. Ethidium bromide saturates the nucleic acids and reduces the efficiency of RNA transfer.
Typical procedure:1. Soak the dried filter in 5% acetic acid at room
temperature for 15 minutes.2. Place the filter in 0.5 M sodium acetate (pH 5.2)
and 0.04% methylene blue for 5-10 minutes at room temperature.
3. Rinse the filter in water for 5-10 minutes.CAS #99-76-3
19220 100 gm 500 gm
N,N’-Methylene-bis-Acrylamide(bis-Acrylamide)
ULTRAPURE
MB Grade(CH2=CHCONH)2CH2
Formula Weight: 154.17Product specifications:Form: White crystalline powderAssay: ≥ 99.9%Identity: By IRA (290 nm, 1%, 1 cm): ≤ 0.2pH (1%, H2O): ≥ 5.5Acrylic acid: ≤ 0.02%Lead (Pb): ≤ 1 ppmGel Performance: Passes testApplication:� Cross-linking agent used in forming polyacryla-
mide gels. The porosity of the gel is determined by the chain length and the degree of cross-linking. Most sequencing gels use a ratio of 1 molecule of cross-linker for every 19 monomers of acrylamide.
CAS #110-26-9
75821 25 gm 50 gm 100 gm 1 kg
5-Methyl-2’-Deoxycytidine-5’-Monophosphate, Disodium Salt(5-Methyl dCMP)
C10H14N3O7PNa2
Formula Weight: 365.19 (anhydrous)Product specifications:Form: White powderAssay (HPLC, as nucleotide): ≥ 98%Assay (HPLC, as nucleoside): ≥ 98%Moisture (KF): ≤ 16%Phosphorylated Nucleosides:≤ 0.20% dCMP≤ 0.049% for any other phosphorylated nucleosideStorage: -20°C, protect from moisture.
19184 100 mg
4-Methylumbelliferyl-α-L-Iduronide, Sodium Salt
C16H15NaO9
Formula Weight: 374.27Storage: -20°C
33230 5 mg
Mineral OilULTRAPURE
MB GradeProduct specifications:Form: Clear, colorless oilIdentity: By IRSpecific Gravity (25°C): 0.818 to 0.880Viscosity (40°C): ≤ 33.5 centistokesNeutrality: Passes testReadily Carbonizable Substances: Passes testLimit of Polynuclear Compounds: Passes testSolid Paraffin: Passes testUse Test: Functionally tested for use in PCR.CAS #8042-47-5
71600 10 ml 25 ml 1 L
Monosodium Glutamate
See Glutamic Acid, Monosodium Salt, page 218
MOPS(3-[N-Morpholino]propanesulfonic Acid)
ULTRAPURE
HO3S(CH2)3(C4H8NO)Formula Weight: 209.26Product specifications:Form: White/clear crystalline powderAssay: ≥ 98.5% (dry basis)Identity: By IRpKa (20°C): 7.20 ± 0.15A (250 nm, 1 M, 1 cm): ≤ 0.1Loss on Drying: ≤ 0.5%Residue on Ignition: ≤ 0.1%Chloride (Cl): ≤ 0.01%Sulfate (SO4): ≤ 0.02%Iron (Fe): ≤ 5 ppmLead (Pb): ≤ 10 ppmCAS #1132-61-2
19256 100 gm 1 kg
MTT(3-[4,5-Dimethyl-2-Thiazolyl]-2,5-Diphenyl Tetrazolium Bromide; Thiazolyl Blue Tetrazolium Bromide)
ULTRAPURE
C18H16BrN5SFormula Weight: 414.32Product specifications:Form: Yellow powderChromatography (TLC): HomogeneousIdentity: By IRSolubility: Passes testSensitivity: Passes testMelting Point: ≥ 192°CApplications:� Used in the quantitation of DNA synthesis.� Detects living and growing cells. MTT is cleaved
by active mitochondria to form a dark blue formazan product.
CAS #298-93-1
19265 500 mg 1 gm 5 gm 10 gm
β-NAD
See Nicotinamide Adenine Dinucleotide, page 229
Biochemicals
229For bulk or alternate pack sizes, email us at [email protected].
NADH
See β-Nicotinamide Adenine Dinucleotide, Reduced, page 230
NADP
See Nicotinamide Adenine Dinucleotide Phosphate, page 230
NADPH
See Nicotinamide Adenine Dinucleotide Phosphate, Reduced, page 230
α-Naphthyl Phosphate, Monosodium Salt
C10H8NaO4P•H2OFormula Weight: 264.15 (monohydrate)Product specifications:Form: White crystalline powderAssay: ≥ 97% (as is monohydrate)Chromatography (TLC): HomogeneousIdentity: By IRMoisture (KF): ≤ 10.0%Storage: 4°CCAS #1136-89-6
19391 25 gm
NBT
See p-Nitro Blue Tetrazolium Chloride, page 230
Neocuproine, Hydrochloride(2,9-Dimethyl-1,10-Phenanthroline HCl)
C14H12N2•HCl•nH2O
Formula Weight: 244.72•nH2OProduct specifications:Form: Off-white to pale greenish-tan powderAssay: ≥ 98.0% (dry basis)Identity: By IRChromatography (TLC): HomogeneousMoisture (KF): ≤ 10%Storage: 4°CCAS #7296-20-0
19430 10 gm 25 gm 100 gm 1 kg
Neomycin SulfateULTRAPURE
USBioAnalyzedC23H46N6O13
•3H2SO4
Formula Weight: 908.88Product specifications:Form: White to off-white powderActivity: ≥ 600 µg neomycin/mg powder (dry basis)Activity: ≥ 552 µg neomycin/mg powder (as is)Identity (A, B, C): Passes testspH (33 mg/ml): 5.0 - 7.5Loss on Drying: ≤ 8.0%Mode of Action: Binds to ribosomal components
and inhibits protein synthesis.Stock Solution: 10 mg/ml in waterWorking Concentration: 10 µg/mlStorage: Ambient, in tightly closed, light-resistant
containers.CAS #1405-10-3
19435 10 gm 25 gm
Neutral Red
C15H17N4ClFormula Weight: 288.78Product specifications:Form: Green-black crystalline powderIdentity: By IRVisual Transition Interval: pH 6.8 - Red pH 8.0 - AmberCAS #553-24-2
19465 25 gm 100 gm 1 kg
Niacinamide(Nicotinic Acid Amide; Nicotinamide)
USBioAnalyzedC6H6N2OFormula Weight: 122.12Product specifications:Form: White crystalline powderAssay: 98.5 - 101.5% (dry basis)Identity (A, B): Passes testsMelting Point: 128 - 131°CLoss on Drying: ≤ 0.5%Residue on Ignition: ≤ 0.1%Heavy Metals: ≤ 0.003%Readily Carbonizable Substances: Passes testResidual Solvents: Meets requirementsCAS #98-92-0
19480 1 kg 10 kg
Nicotinamide
See Niacinamide, page 229
β-Nicotinamide Adenine Dinucleotide(β-NAD; Coenzyme I; Diphosphopyridine Nucleotide; DPN)
C21H27N7O14P2•3H2O
Formula Weight: 717.47 (trihydrate) 663.43 (anhydrous)Product specifications:Form: White powderAssay (Enzymatic): 99 ± 4%Assay: 100 ± 3% (UV, 260 nm, pH 7.5)Identity: By IRε340 (pH 10.0): [6.3 ± 0.2] x 103
ε260 (pH 7.5): [18.0 ± 0.5] x 103
Spectral Ratios (pH 7.5): A250/A260: 0.83 ± 0.03 A280/A260: 0.21 ± 0.02Spectral Ratios (pH 10, reduced with ADH): A340/A260: 0.43 ± 0.01Moisture: ≤ 8%Note: The above product is available in manufactur-
ing quantities. Please inquire for bulk pricing.Storage: -20°C, protect from moisture.CAS #53-84-9
15335 1 gm 5 gm 10 gm 25 gm 100 gm
β-Nicotinamide Adenine Dinucleotide, Lithium Salt, Dihydrate(β-NAD)
C21H26N7O14P2Li•2H2OFormula Weight: 705.39 (dihydrate) 663.43 (free acid)Free of LDH. Stable.Storage: -20°C
19438 1 gm 5 gm
Biochemicals
230 888-362-2447 | 216-765-5000 | usb.affymetrix.com
β-Nicotinamide Adenine Dinucleotide, Monosodium Salt(β-NAD)
C21H26N7O14P2Na•2H2OFormula Weight: 721.44 (dihydrate) 685.41 (anhydrous monosodium)Product specifications:Form: White to pale yellow powderAssay (Enzymatic): ≥ 95%Assay (UV 260 nm, pH 7.5): 100 ± 3%Spectral Ratios (pH 7.5): A250/A260: 0.83 ± 0.03 A280/A260: 0.21 ± 0.02Spectral Ratios (pH 10): A340/A260: 0.43 ± 0.01 A260/A340: 2.32 ± 0.06Moisture: ≤ 8%pH (0.4%, H2O): 5.35 - 6.35Sodium (Na): 3.0% ± 0.5%Storage: -20°C, protect from moisture.CAS #20111-18-6
19439 5 gm
β-Nicotinamide Adenine Dinucleotide-Reduced, Disodium, Trihydrate(β-NADH; Coenzyme I-Reduced; Triphosphopyridine Nucleotide-Reduced; DPNH)
C21H27N7O14P2Na2•3H2O
Formula Weight: 763.45 (disodium, trihydrate) 665.25 (anhydrous, NADA)Product specifications:Form: White to cream colored powderAssay (Enzymatic): 99 ± 4%Assay (UV, 260 nm): 99 ± 4%ε260 (10 mM Tris): [14.4 ± 0.5] x 103
ε340 (10 mM Tris): [6.3 ± 0.2] x 103
Spectral Ratios (10 mM Tris): A250/A260 (red): 0.82 ± 0.03 A280/A260 (red): 0.23 ± 0.02 A340/A260 (red): 0.43 ± 0.01Moisture (KF): ≤ 8%Sodium (Na): 6.5 ± 1.5%Note: Do not use unbuffered water for dissolution
since it may cause decomposition of β-NADH.Storage: -20°C, protect from moisture.CAS #606-68-8
15345 1 gm 5 gm 10 gm 25 gm
β-Nicotinamide Adenine Dinucleotide Phosphate, Monosodium, Tetrahydrate(β-NADP; Coenzyme II-Oxidized; Triphosphopyridine Nucleotide-Oxidized; TPN)
C21H27N7O17P3Na•4H2OFormula Weight: 837.45 (monosodium tetrahydrate) 765.39 (anhydrous, NADP•Na)Product specifications:Form: White powderAssay (Enzymatic): ≥ 93%Spectral Ratios (pH 7.5): A250/A260: 0.83 ± 0.03 A280/A260: 0.21 ± 0.02Spectral Ratios (pH 7.5, reduced with G6PDH): A340/A260: 0.43 ± 0.02 A260/A340: 2.33 ± 0.11ε260 (pH 7.5): [18.0 ± 0.8] x 103
ε340 (pH 7.5): [6.2 ± 0.4] x 103
Moisture: ≤ 8.0%Sodium (Na): 3.0 ± 1.5%Note: The above product is available in manufactur-
ing quantities. Please inquire for bulk pricing.Storage: -20°C, protect from moisture.CAS #1184-16-3
22655 100 mg 250 mg 500 mg 1 gm 5 gm
β-Nicotinamide Adenine Dinucleotide Phosphate-Reduced, Tetrasodium, Tetrahydrate(β-NADPH; Coenzyme II-Reduced; Triphosphopyridine Nucleotide-Reduced; TPNH)
C21H26N7O17P3•Na4
•4H2OFormula Weight: 905.41 (tetrasodium, tetrahydrate)Product specifications:Form: White to slightly off-white powderAssay (Enzymatic): ≥ 93%ε260 (10 mM Tris): [14.4 ± 0.7] x 103
ε340 (10 mM Tris): [6.2 ± 0.3] x 103
Spectral Ratios (10 mM Tris): A340/A260: 0.43 ± 0.01Moisture: ≤ 8.0%Sodium (Na): 10.0 ± 2.0%Note: The above product is available in manufactur-
ing quantities. Please inquire for bulk pricing.Storage: -20°CCAS #2646-71-1
22650 25 mg 100 mg 250 mg 500 mg 1 gm
p-Nitro Blue Tetrazolium Chloride(NBT)
ULTRAPURE
C40H30Cl2N10O6
Formula Weight: 817.64Product specifications:Form: Yellow crystalline powderIdentity: By IRSolubility (0.5% in H2O): Passes testSolubility (1% in MeOH): Passes testSensitivity: Passes testApplication:� Used with 5-bromo-4-chloro-3-indolyl phosphate
(BCIP) to detect antigen-antibody alkaline phos-phatase complexes. The NBT/BCIP mix is convert-ed to a dense blue compound and will develop at the site of antigen-antibody complexes.
NBT Stock Solution: 50 mg/ml in 70% dimethylformamideBCIP/NBT Developing Solution: 33 ml NBT (50 mg/ml) 5 ml AP Buffer (100 mM Tris-HCl, pH 9.5,
100 mM NaCl, 5 mM MgCl2) Add 16.8 ml BCIP stock (50 mg/ml in 100%
dimethylformamide) Protect from light and use within 1 hour.CAS #298-83-9
19535 250 mg 1 gm 5 gm 10 gm
4-Nitrophenyl-N-Acetyl-β-D-Galactosaminide(p-Nitrophenyl-2-Acetamido-2-Deoxy-β-D-Galactoside)
C14H18N2O8
Formula Weight: 342.30Product specifications:Form: White to off-white powderPurity (TLC): ≥ 98%Storage: -20°CCAS #14948-96-0
34128 100 mg 1 gm
Biochemicals
231For bulk or alternate pack sizes, email us at [email protected].
4-Nitrophenyl-α-L-Fucopyranoside
C12H15NO7
Formula Weight: 285.25Product specifications:Form: White powderIdentity: By IRMelting Point: 196 ± 2°CStorage: 4°CCAS #10231-84-2
34131 50 mg 250 mg 1 gm
4-Nitrophenyl-α-D-Galactopyranoside(p-Nitrophenyl-α-D-Galactoside)
C12H15NO8
Formula Weight: 301.25Form: Slightly off-white to yellowish crystalline
powderStorage: -20°CCAS #7493-95-0
34133 500 mg 1 gm 5 gm
4-Nitrophenyl-β-D-Glucopyranoside
C12H15NO8
Formula Weight: 301.25Product specifications:Form: White crystalline powderIdentity: By IR[α]20
D : -106° ± 5° (dry basis, c=1 in H2O)A (425 nm, 8 mg/ml, 1 cm): ≤ 0.07 (in 0.15 M AMP
buffer, pH 10.2)Moisture (KF): ≤ 10%Free p-Nitrophenol: ≤ 0.01%Storage: -20°CCAS #2492-87-7
19594 1 gm 5 gm
4-Nitrophenyl-β-D-Glucuronide
C12H13NO9
Formula Weight: 315.23Storage: 4°CCAS #10344-94-2
34136 100 mg 250 mg 1 gm
4-Nitrophenyl-α-D-Maltoside
C18H25NO13
Formula Weight: 463.39Product specifications:Form: White powderIdentity: By IRPurity (HPLC): ≥ 99%Moisture: ≤ 5.0%Storage: -20°CCAS #17400-77-0
34138 50 mg 100 mg
4-Nitrophenyl-β-D-Mannopyranoside(p-Nitrophenyl-β-D-Mannoside)
C12H15NO8
Formula Weight: 301.25Product specifications:Form: White to off-white crystalline powderIdentity: By IR[α]20
D : -105.0° ± 2° (dry basis, c=4 in H2O)Moisture (KF): ≤ 1.0%Free p-Nitrophenol: ≤ 0.01%Storage: -20°CCAS #35599-02-1
34140 100 mg 500 mg
4-Nitrophenyl-β-D-Xylopyranoside
C11H13NO7
Formula Weight: 271.22Form: White powderStorage: -20°CCAS #2001-96-9
34142 500 mg 1 gm
Nucleoside-5’-Triphosphates (ATP, CTP, GTP, UTP), Set of Four, 100 mM Solutions(NTPs)
ULTRAPURE
100 mM aqueous solutions, pH 7.5Storage: -20°C
4 x 25 µmol (250 µl) each NTP per pack.77245 1 pk
Nucleotide Solutions
See Adenosine-5’-Triphosphate (ATP), page 189See 2’-Deoxyadenosine-5’-Triphosphate (dATP),
page 210See 2’-Deoxycytidine-5’-Triphosphate (dCTP),
page 210See 2’-Deoxyguanosine-5’-Triphosphate (dGTP),
page 210See 2’-Deoxynucleoside-5’-Triphosphates (dNTP),
Set of Four, page 211See 2’-Deoxythymidine-5’-Triphosphate (dTTP),
page 211See 2’-Deoxyuridine-5’-Triphosphate (dUTP),
page 211See 2’,3’-Dideoxyadenosine-5’-Triphosphate
(ddATP), page 212See 2’,3’-Dideoxycytidine-5’-Triphosphate (ddCTP),
page 212See 2’,3’-Dideoxythymidine-5’-Triphosphate
(ddTTP), page 212See Guanosine-5’-Triphosphate (GTP), page 221See PCR-Nucleotide Mixes, page 232
NZCYM Broth, Ready-Made Powder
Form: Off-white free flowing powderpH (2.2%, H2O): 7.0 ± 0.2Contents: Casein Peptone: 10 gm/L Casamino Acids: 1 gm/L Magnesium Sulfate: 0.98 gm/L Yeast Extract: 5 gm/L Sodium Chloride: 5 gm/LTo use: Dissolve 22 gm powder per liter of purified
water. Autoclave.Application: � For the propagation and maintenance of lambda
bacteriophage
75855 1 kg
Oligo (dT)12-18 Primer, MW ≅ 4500
MB GradeForm: Supplied in aqueous solutionConcentration: 25 µM ± 2.5 µM2.5 nmol per tube.Storage: -20°C
77405 100 µl (2.5 nmol)
Biochemicals
232 888-362-2447 | 216-765-5000 | usb.affymetrix.com
Oligo p(dT)12-18 Sodium Salt(Oligo dT)
LyophilizedForm: This oligodeoxythymidylate mixture is synthe-
sized using O-cyanoethyl phosphoramidite chem-istry. After the removal of the protecting groups, the product is converted to the sodium salt and the excess salt removed by gel filtration.
Storage: -20°C
19817 5 units 25 units 100 units
Ovalbumin
See Albumin, Egg, page 199
PABA
See p-Amino Benzoic Acid, page 199
Pancreatin 1X
Source: Porcine PancreasForm: White to off-white powder stabilized with
lactose.Product specifications:Protease: 1.0 - 2.0 x USPAmylase: 1.0 - 2.5 x USPLipase: 2.0 - 10.0 x USP units/mgLoss on Drying: 0.0 - 5.0%Fat: 0.0 - 3.0%Salmonella: NegativeE. coli: NegativeUnit Definition: Pancreatin will convert at least 25
times its weight of potato starch into soluble car-bohydrates in 5 minutes at 40°C and at least 25 times its weight of casein into proteases in
1 hour at 40°C.
19880 200 gm
Paraformaldehyde Solution, 4% in PBS
Description: Paraformaldehyde solution 4% in phos-phate buffered saline
Product specifications:Form: Colorless, clear solutionPurity (Formaldehyde): 91 - 93%pH (25°C): 7.0 - 7.6Storage: 4°C
19943 1 L
PBS, 10X Solution, pH 7.4ULTRAPURE
MB GradeForm: 10X PBS (Phosphate Buffered Saline) solution
consisting of 80.6 mM sodium phosphate, 19.4 mM potassium phosphate, 27 mM KCl, and 1.37 M NaCl in high purity dH2O, pH 7.4. Solution is 0.2 µm filtered and dispensed into durable, square Nalgene bottles. The 5 L size is supplied in an upright carton with a convenient dispensing spout. Dilute to 1X with high purity dH2O for a solution consisting of 10 mM phosphate, pH 7.4 ± 1 2.7 mM potassium chloride and 137 mM NaCl.
pH (25°C, 10X): 7.4 ± 0.1Contaminant Testing: DNase (endo): Passes test DNase (exo): Passes test RNase: Passes test Protease: Passes test
75889 100 ml 1 L 5 L
PCR Nucleotide Mix, 10 mM SolutionULTRAPURE
Form: Premixed dNTPs at 10 mM each dATP, dCTP, dGTP, dTTP.
pH: 7.5Functionally tested in PCR with USB Taq DNA
Polymerase (PN 71160).
77212 500 µl 2 x 500 µl
PCR Nucleotide Mix, 25 mM SolutionULTRAPURE
Form: Premixed dNTPs at 25 mM each dATP, dCTP, dGTP, dTTP.
pH: 7.5Functionally tested in PCR with USB Taq DNA
Polymerase (PN 71160).
77119 500 µl
PCR Nucleotide Mix with dUTP, 10 mM SolutionULTRAPURE
Form: Premixed dNTPs at 10 mM each dATP, dCTP, dGTP, dUTP
pH: 7.5Functionally tested in PCR with USB Taq DNA
Polymerase (PN 71160).
77330 500 µl
PEG
See Polyethylene Glycol 400, page 235See Polyethylene Glycol 6000, page 235See Polyethylene Glycol 8000, page 235
Penicillin G, Potassium
USBioAnalyzedC16H17KN2O4SFormula Weight: 372.48Product specifications:Form: White powderAssay: 1440 - 1680 Penicillin G units/mg (as is)Identity A: By IRIdentity B: Passes testpH (60 mg/ml): 5.0 - 7.5Loss on Drying: ≤ 1.5%Crystallinity: Passes testMode of Action: Interferes with bacterial cell wall synthesis and
cell division. Inactivated by oxidizing and reducing agents,
glycerols and alcohols.CAS #113-98-4
19985 1 kg
Pepsin
(E.C.3.4.23.1)Source: Porcine StomachProduct specifications:Form: Lyophilized, salt free, 2X crystallizedActivity: ≥ 2500 units/mgUnit Definition: One unit causes an increase in
absorbance at 280 nm of 0.001/minute at 37°C, pH 2.0.
Storage: -20°CCAS #9001-75-6
20010 1 gm 5 gm 10 gm 25 gm
Biochemicals
233For bulk or alternate pack sizes, email us at [email protected].
Pepsin 1:10,000
(E.C.3.4.23.1)Source: Porcine StomachProduct specifications:Form: Cream colored powderActivity: 1:10,000 - 11,500Loss on Drying: ≤ 4.0%Storage: Protect from moisture.
20015 100 gm 500 gm 1 kg
Pepstatin A(Isovaleryl-Val-Val-4-Amino-3-Hydroxy-6-Methylheptanoyl-Ala-4-Amino-3-Hydroxy-6-Methylheptanoic Acid)
ULTRAPURE
C34H63N5O9
Formula Weight: 685.89Product specifications:Form: White powderChromatography (TLC): HomogeneousPeptide Content: ≥ 98%Biological Activity: ≥ 850 µg/mgApplication:� Pepstatin A, a protease inhibitor, inhibits Pepsin,
Renin, and Cathepsin D. Effective concentrations are 10-6 to 10-9 M. It does not inhibit Trypsin, Chymotrypsin, Papain, or Plasmin.
Note: Dissolves in methanol.Storage: 4°CCAS #26305-03-3
20037 5 mg 25 mg
Peptones
See Bacteriological Peptone, page 202See Casein Hydrolysate, page 205See Yeast Extract and Hydrolysate, pages 251
Phenol(Carbolic Acid)
ULTRAPURE
MB GradeC6H5OHFormula Weight: 94.11Product specifications:Form: Translucent crystalline solidAssay: ≥ 99%Clarity of Solution (5%, H2O): Passes testpH (5%, H2O): 4.5 - 6.0Freezing Point: ≥ 40.5°CWater: ≤ 0.1%Res. on Evaporation: ≤ 0.05%Buffer Equilibration Suitability: Passes testIron (Fe): ≤ 1 ppmLead (Pb): ≤ 2 ppmMagnesium (Mg): ≤ 1 ppmDNase (endo): Passes testPreservatives/Inhibitors: Passes testApplication:� Used primarily for DNA/RNA extraction and puri-
fication.Preparation of buffered phenol:1. Allow phenol to warm to room temperature and
then melt at 68°C.2. Add an equal volume of buffer (0.5 M Tris-HCl,
pH 8.0).3. Stir for about 15 minutes on a magnetic stirrer.4. When the two phases have separated, aspirate
the top (aqueous) phase.5. Add an equal volume of 0.1 M Tris-HCl, pH 8.0
and repeat stirring.6. Remove the aqueous phase and repeat extrac-
tions until the pH of the phenol is 7.8. (Note: Measure with pH paper).
7. After the phenol is equilibrated, remove the final aqueous phase. Top with 0.1 volumes of 0.1 M Tris-HCl, pH 8.0.
8. Store in a light-tight bottle at 4°C.Note: For convenience, we provide phenol which is
already equilibrated (PN 75829).Storage: -20°CPacked under inert atmosphereCAS #108-95-2
75830 500 gm
Phenol, pH 8.0, EquilibratedULTRAPURE
MB GradeForm: This product is supplied as water-saturated
phenol with a separate bottle of Phenol Equil-ibration Buffer. At the time of product use, simply add the entire contents of Phenol Equilibration Buffer to the water-saturated phenol.
pH (5%, after equilibration, 20°C): ≥ 8.0Contaminant Testing: DNase: Passes testStorage: 4°CApplication: � Used in nucleic acid purification procedures and
dissolution of protein
75829 100 ml 400 ml
Phenol: Chloroform: Isoamyl Alcohol (25:24:1)ULTRAPURE
MB GradeForm: A liquid mixture of phenol, ACS Reagent
Grade chloroform and ACS Reagent Grade iso-amyl alcohol in a ratio of 25:24:1 supplied with an additional Alkaline Equilibration Buffer used to adjust the pH to 8.0.
Note: Phenol:Chloroform:Isoamyl Alcohol (25:24:1) is packaged at pH 6.7. The product is accompa-nied by a separate Alkaline Equilibration Buffer. When ready to use add the supplemental buffer’s entire contents to the Phenol:Chloroform:Isoamyl Alcohol, then mix or shake, and allow to separate. Once this is completed the product will be at a pH of 8.0.
Contaminant Testing: DNase: Passes testpH (5%, H2O): 8.0Application: � Nucleic acid extraction and purificationPhenol/Chloroform RNA Isolation MethodA. Guanidine HCl Homogenization Buffer for RNA
Isolation: 8.0 M Guanidine HCl, 0.1 M Sodium Acetate, pH 5.5, 5.0 mM Dithiothreitol, 0.5% Sodium Lauryl Sarcosinate.
B. Extraction with Phenol: Most extractions are per-formed using equal volumes of equilibrated phe-nol with respect to the sample volume. Normally, the extraction is performed twice with phenol and once with a 1:1 solution of phenol/chloro-form. Hence, it is estimated that 25 ml of phenol is required for every 10 ml of sample.
Note: If regular phenol is purchased, it will need to be equilibrated before use. For this reason, many customers may prefer to purchase phenol that is already equilibrated (PN 75829).
C. RNA Resuspension Buffer: TE, pH 7.5.Storage: 4°C
75831 100 ml 400 ml
Biochemicals
234 888-362-2447 | 216-765-5000 | usb.affymetrix.com
Phenolphthalein
ACS Reagent GradeC20H14O4
Formula Weight: 318.32Product specifications:Form: White to off-white powderClarity of Alcohol Solution (1%, EtOH): Passes testVisual Transition: From pH 8.0: Colorless to pH 10.0: RedCAS #77-09-8
20075 500 gm 1 kg
Phenol Red, Sodium Salt(Phenolsulfonphthalein)
C19H13O5SNaFormula Weight: 376.36Product specifications:Form: Rust/red crystalline powderIdentity: By IRClarity of Solution: Passes testVisual Transition Interval: pH 6.8 - Yellow pH 8.2 - RedCAS #34487-61-1
20085 10 gm 25 gm 50 gm
Phenolsulfonphthalein
See Phenol Red, page 234
DL-Phenylalanine
C6H5CH2CH(NH2)COOHFormula Weight: 165.19Product specifications:Form: White crystalline powderAssay: ≥ 99.0%Chromatagraphy (TLC): HomogeneousIdentity: By IRLoss on Drying: ≤ 0.3%Residue on Ignition: ≤ 0.3%Heavy Metals (Pb): ≤ 10 ppmAmmonia (NH3): ≤ 0.03%Arsenic (As): ≤ 3 ppmChloride (Cl): ≤ 0.02%Iron (Fe): ≤ 50 ppmCAS #150-30-1
20100 100 gm 1 kg
L-Phenylalanine
USBioAnalyzed(C6H5)CH2CH(NH2)COOHFormula Weight: 165.19Product specifications:Form: White crystalline powderAssay: 98.5 - 101.5% (dry basis)Chromatographic Purity: Passes testIdentity: By IRLoss on Drying: ≤ 0.3%[α]25
D : -32.7° to -34.7° (c=2 in H2O)pH (1%, H2O): 5.4 - 6.0Residue on Ignition: ≤ 0.4%Sulfate (SO4): ≤ 0.03%Heavy Metals (Pb): ≤ 0.0015%Chloride (Cl): ≤ 0.05%Iron (Fe): ≤ 0.003%Residual Solvents: Meets requirementsCAS #63-91-2
20105 100 gm 250 gm 1 kg 5 kg
Phenylmethyl Sulfonyl Fluoride(PMSF)
ULTRAPURE
C7H7FO2SFormula Weight: 174.19Product specifications:Form: Off-white/clear crystalsAssay: ≥ 99.0%Identity: By IRApplication: � PMSF is a protease inhibitor active against trypsin
and chymotrypsin.CAS #329-98-6
20203 5 gm 25 gm 100 gm
Phosphocreatine
See Creatine Phosphate, page 208
PIPES(Piperazine-N,N'-bis-[2-Ethanesulfonic Acid])
C8H18N2O6S2
Formula Weight: 302.37Product specifications:Form: White/clear crystalline powderAssay: ≥ 98.5% (dry basis)Identity: By IRpKa (20°C): 6.80 ± 0.15A (250 nm, 5% in 1N NaOH, 1 cm): ≤ 0.20Loss on Drying: ≤ 0.5%Iron (Fe): ≤ 30 ppmHeavy Metals (Pb): ≤ 10 ppmCAS #5625-37-6
20426 100 gm 1 kg
PIPES, Disodium Salt
C8H16N2O6S2Na2
Formula Weight: 346.33Product specifications:Form: White powderAssay: ≥ 98.0% (dry basis)Identity: By IRpKa (20°C): 6.80 ± 0.15A (250 nm, 0.1 M, 1 cm): ≤ 0.1Loss on Drying: ≤ 10.0%CAS #76836-02-7
20428 10 gm 100 gm 1 kg
PMSF
See Phenylmethyl Sulfonyl Fluoride, page 234
Polyanetholesulfonic Acid, Sodium Salt
(C10H11NaO4S)n
Formula Weight: (250.25)n
CAS #55963-78-5
20450 5 gm 100 gm
Biochemicals
235For bulk or alternate pack sizes, email us at [email protected].
Poly(dI-dC) • Poly(dI-dC) Sodium Salt
Product specifications:Description: Lyophilized, sodium salt complex of two
strands, each with an alternating sequence of deoxyinosinic acid and deoxycytidylic acid.
Form: White to off-white lyophilized powderConcentration: One A260 unit of double-stranded
polymers is ~ 50 µg.UV Absorption Spectrum: λmax: 248-252 nmAbsorbance ratios: A250/A260: 1.15-1.31 A280/A260: 0.64-0.74All UV Absorbance data were obtained in 0.1 M
sodium chloride, 0.02 M potassium phosphate, pH 7.0.
Note: To maintain a double-stranded configura-tion of the polymer in solution, rehydrate with a neutral buffer containing 100 mM NaCl. After reconstitution store the solution at -20°C.
Storage:-20°C
20539 5 units 25 units 100 units
Polyethylene Glycol 400(PEG 400)
Formula Weight: 380 - 420Product specifications:Form: Clear colorless viscous liquidDensity (d20
4 ): 1.12 - 1.13Refractive Index (n20
d ): 1.465 - 1.469Viscosity (20°C): 102 - 120 mpasCAS #25322-68-3
19957 500 ml 1 L
Visit our website, usb.affymetrix.com, for the latest product information.
Polyethylene Glycol 6000, Flakes(PEG 6000)
Formula Weight: 5400 - 6600Product specifications:Form: White or slightly yellowish flakesSolubility (50%, Hot H2O): Clear and colorlessMelting Point: Approx. 60°CApplications:� Differential precipitation of DNA� Enhances hybridization rate of nucleic acids.� Improves efficiency of end-labeling with T4
Polynucleotide Kinase(1,2).� Used in ligation of blunt ended DNA.� Precipitates bacteriophage ± particles and plas-
mid DNA.� Used in preparing single-stranded M13 DNA.� Promotes cell fusion.PEG, with magnesium, causes DNA to undergo a
“psi” transition and it collapses into a highly condensed state(1). This results in macromolecular crowding(2) and increased efficiency in end-labeling.
References:1. Lerman, L. S. (1971) Proc Natl. Acad Sci, 68, 1886.2. Harrison, B. and Zimmerman, S. B., (1986) Anal
Biochem, 158, 307.CAS #25322-68-3
19972 1 kg 5 kg
Polyethylene Glycol 8000, Flakes(PEG 8000)
ULTRAPURE
Approx. M.W.=8,000Product specifications:Form: White flakesSolubility: Clear and colorlessMelting Point: 60 - 64°CApplications:� Differential precipitation of DNA� Enhances hybridization rate of nucleic acids.� Improves efficiency of end-labeling with T4
Polynucleotide Kinase(1,2).� Used in ligation of blunt ended DNA.� Precipitates bacteriophage ± particles and plas-
mid DNA.� Used in preparing single-stranded M13 DNA.� Promotes cell fusion.PEG, with magnesium, causes DNA to undergo a
“psi” transition and it collapses into a highly condensed state(1). This results in macromolecular crowding(2) and increased efficiency in end-labeling and blunt ligation.
References:1. Lerman, L. S. (1971) Proc Natl. Acad Sci, 68, 1886.2. Harrison, B. and Zimmerman, S. B., (1986) Anal
Biochem, 158, 307-315.CAS #25322-68-3
19959 500 gm 1 kg 5 kg
Polyethylene Glycol 8000, Powder(PEG 8000)
Molecular Weight Range: 7,000 - 9,000Product specifications:Form: White to off-white powderSolubility (50%, hot H2O): Clear and colorlessMelting Point: 60 - 64°CApplications:� Differential precipitation of DNA� Enhances hybridization rate of nucleic acids.� Improves efficiency of end-labeling with T4
Polynucleotide Kinase(1,2).� Used in ligation of blunt ended DNA.� Precipitates bacteriophage ± particles and plas-
mid DNA.� Used in preparing single-stranded M13 DNA.� Promotes cell fusion.PEG, with magnesium, causes DNA to undergo a
“psi” transition and it collapses into a highly condensed state(1). This results in macromolecular crowding(2) and increased efficiency in end-labeling and blunt ligation.
References:1. Lerman, L. S. (1971) Proc Natl. Acad Sci, 68, 1886.2. Harrison, B. and Zimmerman, S. B., (1986) Anal
Biochem, 158, 307.CAS #25322-68-3
19966 500 gm 1 kg 5 kg
Polymixin B Sulfate
USBioAnalyzedProduct specifications:Form: White to buff colored powderActivity: ≥ 6000 Polymixin B units/mg (dry basis) ≥ 5580 Polymixin B units/mg (as is)Identity (A, B, C): Passes testspH (5 mg/ml): 5.0 - 7.5Loss on Drying: ≤ 7.0%Content of Phenylalanine: 9%-12%CAS #1405-20-5
20551 10 mu 100 mu
Polyvinyl Pyrrolidone (PVP)
See PVP-K-30, page 237
Biochemicals
236 888-362-2447 | 216-765-5000 | usb.affymetrix.com
Ponceau S, Sodium SaltULTRAPURE
(C.I.27195)C22H12N4O13S4Na4
Formula Weight: 760.57Application: � As a protein stain for serum proteins separated by
agarose gel and cellulose acetate electrophoresisCAS #6226-79-5
32819 50 gm
Potassium ChlorideULTRAPURE
ACS Reagent GradeKClFormula Weight: 74.55Product specifications:Form: White crystalline powderAssay: 99.0 - 100.5%pH (5%, H2O, 25°C): 5.4 - 8.6Insoluble Matter: ≤ 0.005%Iodide (I): ≤ 0.002%Bromide (Br): ≤ 0.01%Chlorate & Nitrate (NO3): ≤ 0.003%Nitrogen Cmpds. (N): ≤ 0.001%Phosphate (PO4): ≤ 5 ppmSulfate (SO4): ≤ 0.001%Barium (Ba):Passes testHeavy Metals (Pb): ≤ 5 ppmIron (Fe): ≤ 3 ppmCalcium (Ca): ≤ 0.002%Magnesium (Mg): ≤ 0.001%Sodium (Na): ≤ 0.005%CAS #7447-40-7
20598 500 gm 1 kg 5 kg
Potassium Chloride (KCl), 2 M SolutionULTRAPURE
MB GradeForm: 2 M solution of potassium chloride (KCl) in
high purity dH2O. Solution is 0.2 µm filtered and dispensed into durable, square Nalgene bottles.
Contaminant Testing: DNase (endo): Passes test DNase (exo): Passes test RNase: Passes testCAS #7447-40-7
75896 100 ml
Potassium Phosphate, DibasicULTRAPURE
ACS Reagent GradeK2HPO4
Formula Weight: 174.18Product specifications:Form: White crystalline powderAssay: ≥ 98.5%pH (5%, H2O, 25°C): 8.5 - 9.6Insoluble Matter: ≤ 0.01%Loss on Drying: ≤ 1.0%Chloride (Cl): ≤ 0.003%Nitrogen Compds. (N): ≤ 0.001%Sulfate (SO4): ≤ 0.005%Heavy Metals (Pb): ≤ 5 ppmIron (Fe): ≤ 0.001%Sodium (Na): ≤ 0.05%Storage: Protect from moisture (product is deliques-
cent).CAS #7758-11-4
20274 500 gm 1 kg 2.5 kg
Potassium Phosphate, Dibasic(Dipotassium Phosphate)
CP GradeK2HPO4
Formula Weight: 174.18Product specifications:Form: White deliquescent powderAssay: ≥ 98.0% K2HPO4 (dry basis)Identity: By IRArsenic (As): ≤ 3 ppmFluoride (F): ≤ 10 ppmInsol. Substances: ≤ 0.2%Lead (Pb): ≤ 2 ppmLoss on Drying: ≤ 2.0%Note: This product is a general laboratory reagent
suitable for most laboratory animal diets where trace element deficiencies are not required. One molar stock solutions of this product may be slightly hazy. Where use of a high quality research buffer is required, we recommend the use of our ACS Reagent Grade phosphate buffers.
Storage: Dessicate (product is deliquescent).CAS #7758-11-4
20276 1 kg 10 kg
Potassium Phosphate, Monobasic, AnhydrousULTRAPURE
ACS Reagent GradeKH2PO4
Formula Weight: 136.09Product specifications:Form: White crystalline powderAssay: ≥ 99.0%pH (5%, H2O, 25°C): 4.1 - 4.5Insoluble Matter: ≤ 0.01%Loss on Drying: ≤ 0.2%Chloride (Cl): ≤ 0.001%Nitrogen Compds. (N): ≤ 0.001%Sulfate (SO4): ≤ 0.003%Heavy Metals (Pb): ≤ 0.001%Iron (Fe): ≤ 0.002%Sodium (Na): ≤ 0.005%CAS #7778-77-0
20227 500 gm 1 kg
Potassium Phosphate, Monobasic
KH2PO4
Formula Weight: 136.09Product specifications:Form: White crystalline powderAssay: ≥ 98.0% (dry basis)Arsenic (As): ≤ 3 ppmFluoride (F): ≤ 10 ppmInsoluble Substances: ≤ 0.2%Lead (Pb): ≤ 2 ppmLoss on Drying: ≤ 1%Note: This product is a general laboratory reagent
suitable for most laboratory animal diets where trace element deficiencies are not required. One molar stock solutions of this product may be slightly hazy. Where use of a high quality research buffer is required, we recommend the use of our ACS Reagent Grade phosphate buffers.
CAS #7778-77-0
20228 1 kg 10 kg
Biochemicals
237For bulk or alternate pack sizes, email us at [email protected].
Protamine Sulfate
Source: SalmonProduct specifications:Form: White to off-white powderAssay: Each mg neutralizes not less than 100 USP
Heparin Units (dry basis)Loss on Drying: ≤ 5.0%Nitrogen (N): 22.5 - 25.5% (dry basis)Sulfate (SO4): 16.0 - 22.0% (dry basis)CAS #9009-65-8
20795 25 gm 100 gm
Proteinase K
Source: Tritirachium albumProduct specifications:Form: Lyophilized powderConcentration: 20 - 50 units/mgMolecular Weight: 28.9 kDaUnit Definition: One unit is the amount of enzyme
that liberates folin positive amino acids and peptides corresponding to 1 µmol tyrosine in 1 minute at 37°C using hemoglobin as substrate.
Application: � Proteinase K inactivates DNase and RNase from
most mammalian cells and microorganisms in the presence of 0.5 - 1% SDS. It can also be used for research involving protein structure because of its cleavage specificity.
Storage: Shipped on cold pack. Store dry at 4°C.CAS #39450-01-6
76230Y 100 mg76230Z 500 mg76230 1 gm (2 x 500 mg)
pUC 19 DNA
See Molecular Biology Products & Kits section, page 129
Purification Solutions
Phenol, pH 8.0, Equilibrated, page 233Phenol:Chloroform:Isoamyl Alcohol (25:24:1),
page 233
PVP-K-30(Polyvinylpyrrolidone)
Formula Weight: 40,000 (average)Product specifications:Form: Off-white powderActive content: ≥ 95%K-value (viscosity of 1% solids w/v aqueous solu-
tion): 26 - 34Moisture: ≤ 5.0%% Unsaturation (calculated as % vinylpyrrolidone):
≤ 0.5%CAS #9003-39-8
20611 1 kg
Pyruvic Acid, Sodium Salt(Sodium Pyruvate)
CH3COCOONaFormula Weight: 110.04Product specifications:Form: White to slightly yellow crystalline powderAssay: ≥ 98.5%Identity: By IRMoisture (KF): ≤ 1.0%Chloride (Cl): ≤ 30 ppmSulfate (SO4): ≤ 0.01%Heavy Metals (Pb): ≤ 10 ppmCAS #113-24-6
20995 10 gm 25 gm 100 gm 500 gm
D-(+)-Raffinose, Pentahydrate
C18H32O16•5H2O
Formula Weight: 594.51Product specifications:Form: White crystalline powderIdentity: By IR[α]20
D : +104.9° ± 1.3° (c=4 in H2O & NH3) (cor-rected to a theoretical pentahydrate)
Loss on Drying: ≤ 15.3%Residue on Ignition: ≤ 0.05%Iron (Fe): ≤ 5 ppmHeavy Metals (Cu): ≤ 10 ppmArsenic (As): ≤ 0.5 ppmCAS #17629-30-0
21060 100 gm 500 gm 1 kg
RapidGel 40% Liquid Acrylamide Stock SolutionULTRAPURE
MB GradeForm: Pre-mixed, 40% liquid acrylamide (19:1
acrylamide:bis-acrylamide ratio) in ready-to-use form for Polyacrylamide Gel Electrophoresis (PAGE). Stock solution provides flexibility for dif-ferent acrylamide percentages in a wide variety of gel applications. This formulation does not contain urea.
Testing: Functionally tested in gel electrophoresis.To use: Dilute 40% stock solution to desired con-
centration with appropriate running buffer for PAGE. To make 100 ml of a sequencing gel solu-tion containing 7-8.3 M urea, add an appropriate volume of 40% RapidGel solution, 10 ml of 10X (or 5 ml of 20X) running buffer, 42-50 gm of urea, and high purity dH2O to 100 ml in a clean, graduated cylinder. Stir and warm solution at 35-45°C to dissolve urea. (Do not microwave.) Cool to room temperature. Filter through a 0.2 µm filter and leave under gentle vacuum for 10 minutes to de-gas. Add 600 µl 10% ammoni-um persulfate and 60 µl TEMED per 100 ml gel solution to polymerize. Allow to polymerize at room temperature from one hour to overnight. For polymerization within 30-45 minutes, add 1 ml 10% ammonium persulfate and 100 µl TEMED per 100 ml gel solution. Pour gel immediately.
Storage: 4°C
75848 500 ml
Biochemicals
238 888-362-2447 | 216-765-5000 | usb.affymetrix.com
RapidGel-XL 6% Liquid Acrylamide DNA Sequencing Gel, TBE FormulationULTRAPURE
MB GradeForm: Pre-mixed, liquid acrylamide in ready-to-
use form for Ultrapure DNA sequencing gels. Contains TBE Buffer and 7 M urea. The most commonly requested gel formulation, RapidGel-XL, 6%, is a high resolution matrix for DNA sequenc-ing, yielding up to 40% more readable sequence per gel when using either radioactive or fluores-cent labeling methods. No gel fixing is necessary.
Testing: Functionally tested in sequencing gel elec-trophoresis.
To use: Intended for use with 1X TBE Buffer. De-gas solution by placing under gentle vacuum for 10 minutes at room temperature. Add 600 µl 10% ammonium persulfate and 60 µl TEMED per 100 ml gel solution to polymerize. Allow to polymerize at room temperature from one hour to overnight. For polymerization within 30-45 minutes, add 1 ml 10% ammonium persul-fate and 100 µl TEMED per 100 ml gel solution. Pour gel immediately. Do not fix gel.
Note: Urea may crystallize out of solution when the gel mix is stored as recommended at 4°C. This is normal and will not affect performance. If this occurs, warm the gel mix at 37°C for 10 minutes or until the urea redissolves. Mix gently. Cool to room temperature before adding the ammonium persulfate and TEMED.
Use with corresponding TBE Running Buffer: 10X Ready Mixed Powder, PN 70454, packaged in 6 x 200 ml bottles or TBE Buffer, 5X Solution, PN 75891, packaged in 1 and 5 liter sizes.
Storage: 4°C
75861 500 ml
RapidRun Agarose Buffer, 20X SolutionULTRAPURE
MB GradeForm: RapidRun Agarose Buffer allows for gels to be
run in one-fourth the time required for standard buffers, such as TAE and TBE. The buffer is a non-borate weak acid that enables high voltage and more rapid electrophoretic runs. Temperature seldom exceeds 38°C during high voltage runs, minimizing the band diffusion and gel degrada-tion commonly associated with TAE and TBE under the same high voltage conditions.
20X RapidRun Agarose Buffer consists of 1.78 M Tris, 0.57 M Taurine, and 0.01 M EDTA. The solu-tion is 0.2 µm filtered.
pH (1:20, 25°C): 8.9 + 0.1Conductivity (1X): 830 + 100 µmhos/cmTesting: Meets or exceeds functional test criteria for
high voltage (300v, 15 minutes) mini-agarose gel electrophoresis.
Contaminant Testing: DNase (endo): Passes test DNase (exo): Passes test RNase: Passes test
Applications:� High voltage, fast agarose gel electrophoresis
(20-30 volts/cm).� This buffer is compatible with commercial gel
extraction kits. However, the pH should be adjust-ed with sodium acetate, pH 5.0, as recommended by the manufacturer.
Note: 1 ml of RapidRun Loading Dye (PN 77524) is included with each pack size. The dye front migrates lower than 50 bases.
Storage: Ambient. Crystals which form at low tem-peratures can be re-dissolved by warming briefly.
77523 1 L 5 L
RapidRun Loading DyeULTRAPURE
MB GradeForm: 5X loading dye (0.05% dye in 50% glycerol).Contaminant Testing: DNase (endo): Passes test DNase (exo): Passes test RNase: Passes testApplication: � Used as a loading dye designed for high voltage
agarose gel electrophoresis with buffers such as RapidRun Agarose Buffer (PN 77523). It may also be used for standard voltage gel electro-phoresis. RapidRun Loading Dye develops an intense orange color in the presence of RapidRun Agarose Buffer. The dye front migrates lower than 50 bases.
77524 1 ml 5 ml
Riboflavin(Vitamin B2 ; Vitamin G)
USBioAnalyzedC17H20N4O6
Formula Weight: 376.36Product specifications:Form: Yellow-orange crystalline powderAssay: 98.0 - 102.0% (dry basis)Identity: By IR[α]25
D : -115° to -135° (c=0.5 in 0.05 M NaOH)Loss on Drying: ≤ 1.5%Residue on Ignition: ≤ 0.3%Limit of Lumiflavin: Passes testResidual Solvents: Meets requirementsStorage: Protect from light.CAS #83-88-5
21145 25 gm 100 gm 500 gm 1 kg
D-(–)-Ribose
C5H10O5
Formula Weight: 150.13Product specifications:Form: White powderAssay: ≥ 98.0% (dry basis)[α]20
D : -20.4° ± 0.4° (c=2 in H2O, NH3)Loss on Drying: ≤ 0.5%Residue on Ignition: ≤ 0.1%Arsenic (As): ≤ 0.5 ppmIron (Fe): ≤ 5 ppmHeavy Metals (Cu): ≤ 10 ppmCAS #50-69-1
21220 10 gm 100 gm
Rifampin
C43H58N4O12
Formula Weight: 822.94Product specifications:Form: Reddish brown crystalline powderAssay: 95.0 - 103.0% of C43H58N4O12 (dry basis) 93.0 - 103.0% of C43H58N4O12 (as is)Identity: By IRpH (1%, H2O): 4.5 - 6.5Loss on Drying: ≤ 2.0%Related Substances: Passes testMode of Action: Inhibits RNA synthesis by binding
to and inhibiting the β subunit of RNA polym-erase.
Storage: Light-resistant containers, protect from heat.
CAS #13292-46-1
21246 1 gm 5 gm
RNA, Free Acid
Source: Torula YeastProduct specifications:Form: White powderMoisture: ≤ 8.0%Nitrogen: ≥ 15.3% (dry basis)Phosphorous: ≥ 9.0% (dry basis)CAS #73049-77-1
21185 100 gm 1 kg
Biochemicals
239For bulk or alternate pack sizes, email us at [email protected].
RNA, Sodium Salt
Source: YeastProduct specifications:Form: White powderMoisture: 4.0 - 8.0%Phosphorous: ≥ 8.0%Nitrogen: ≥ 14.5%pH (1% H2O): 6.5 ± 0.5Biuret test: NegativeCAS #73049-77-1
21190 25 gm 100 gm 1 kg
RNase A, Lyophilized
(E.C. 3.1.4.22)Source: Bovine pancreasForm: Lyophilized powderUnit Definition:One unit is that amount of enzyme
causing the hydrolysis of RNA at a rate such that K equals unity at 25°C and pH 5.0.
Storage: -20°C, desiccatedCAS #9001-99-4
21195 100 mg 1 gm 5 gm
SAMe-PTS
See S-Adenosyl-L-Methionine Sulfate p-Toluenesulfonate, page 190
SDS
See Sodium Dodecyl Sulfate, page 240
DL-Serine (Non-Animal)
CH2OHCHNH2COOHFormula Weight: 105.09Product specifications:Form: White crystalline powderAssay: ≥ 98.5% (dry basis)Solubility (2%, H2O): Clear, colorless solutionIdentity: By IRLoss On Drying: ≤ 0.3%Residue on Ignition: ≤ 0.1%Chloride (Cl): ≤ 0.021%Sulfate (SO4): ≤ 0.05%Heavy Metals (Pb): ≤ 20 ppm
21486 100 gm 1 kg
Sodium Acetate (NaOAc), 3 M Solution, pH 5.5ULTRAPURE
MB GradeForm: 3 M solution of sodium acetate (NaOAc) in
high purity dH2O, pH 5.5. Solution is 0.2 µm fil-tered and dispensed into durable, square Nalgene bottles.
pH: 5.5 ± 0.1Contaminant Testing: DNase (endo): Passes test DNase (exo): Passes test RNase: Passes test
75897 100 ml 500 ml
Sodium Aspartate
See Aspartic Acid, Sodium Salt, page 201
Sodium AzideULTRAPURE
NaN3
Formula Weight: 65.01Product specifications:Form: White crystalline powderAssay: ≥ 99.0%Loss on Drying: ≤ 0.5%CAS #26628-22-8
21610 100 gm 500 gm
Sodium Cacodylate
See Cacodylic Acid, Sodium Salt, page 204
Sodium Carbonate, Anhydrous
ACS Reagent GradeNa2CO3
Formula Weight: 105.99Product specifications:Form: White crystalline powderAssay: ≥ 99.5%Insoluble Matter: ≤ 0.01%Loss on Drying (285°C): ≤ 1.0%Chloride (Cl): ≤ 0.001%Nitrogen Compounds (N): ≤ 0.001%Phosphate (PO4): ≤ 0.001%Silica (SiO4): ≤ 0.005%Sulfur Compounds (SO4): ≤ 0.003%Ammonium Hydroxide Precipitate: ≤ 0.01%Heavy Metals (Pb): ≤ 5 ppmIron (Fe): ≤ 5 ppmCalcium (Ca): ≤ 0.03%Magnesium (Mg): ≤ 0.005%Potassium (K): ≤ 0.005%CAS #497-19-8
21616 500 gm 2.5 kg
Sodium Caseinate
See Casein, Sodium, page 205
Sodium Chloride (NaCl), 5 M SolutionULTRAPURE
MB GradeForm: 5 M solution of sodium chloride (NaCl) in high
purity dH2O. Solution is 0.2 µm filtered and dis-pensed into durable, square Nalgene bottles.
Contaminant Testing: Heavy Metals: ≤ 5 ppm DNase (endo): Passes test DNase (exo): Passes test RNase: Passes test Protease: Passes test
75888 100 ml 500 ml 1 L
Sodium ChlorideULTRAPURE
ACS Reagent GradeNaClFormula Weight: 58.44Product specifications:Form: White/clear crystalline powderAssay: ≥ 99.0%pH (5%, H2O, 25°C): 5.0 - 9.0Insoluble Matter: ≤ 0.005%Iodide (I): ≤ 0.002%Bromide (Br): ≤ 0.01%Chlorate & Nitrate (NO3): ≤ 0.003%Nitrogen Cmpds. (N): ≤ 0.001%Phosphate (PO4): ≤ 5 ppmSulfate (SO4): ≤ 0.004%Barium (Ba): Passes testHeavy Metals (Pb): ≤ 5 ppmIron (Fe): ≤ 2 ppmCalcium (Ca): ≤ 0.002%Magnesium (Mg): ≤ 0.001%Potassium (K): ≤ 0.005%Application: � Used for the precipitation of DNA.CAS #7647-14-5
21618 500 gm 1 kg 5 kg 10 kg
Sodium Citrate
See Citric Acid, Trisodium Salt, page 207
Sodium Deoxycholate
See Deoxycholic Acid, Sodium Salt, page 210
Biochemicals
240 888-362-2447 | 216-765-5000 | usb.affymetrix.com
Sodium Dodecyl Sulfate (SDS)(Sodium Lauryl Sulfate)
ULTRAPURE
CH3(CH2)11OSO3NaFormula Weight: 288.38Product specifications:Form: White crystalline flakesAssay: ≥ 99% (acidity)Assay: ≥ 98% (GC)A (280 nm, 3%, 1 cm): ≤ 0.1A (230 nm, 3%, 1 cm): ≤ 0.2Loss on Drying: ≤ 1%Insoluble Matter: ≤ 0.003%Chloride (Cl): ≤ 0.01%Phosphate (PO4): ≤ 1 ppmCopper (Cu): ≤ 5 ppmIron (Fe): ≤ 1 ppmLead (Pb): ≤ 5 ppmProtocol for 10% SDS Solution: SDS: 100 gm H2O: 900 ml Heat to 68° to dissolve. Adjust to pH 7.2 and
bring to 1 liter. There is no need to sterilize 10% SDS.
CAS #151-21-3
75819 100 gm 1 kg
Sodium Dodecyl Sulfate (SDS)(Sodium Lauryl Sulfate)
CH3(CH2)11OSO3NaFormula Weight: 288.38Product specifications:Form: White crystalline powderIdentity: By IRAssay: ≥ 95% (dry basis)Solubility (20%, H2O): Clear, faintly yellow solutionCAS #151-21-3
18220 500 gm 1 kg 5 kg
Sodium Dodecyl Sulfate (SDS), 10% SolutionULTRAPURE
Form: 10% (w/v) solution of sodium dodecyl sulfate (SDS) in high purity dH2O. Solution is 0.2 µm fil-tered and dispensed into durable, square Nalgene bottles.
A (280 nm, 3%, 1 cm): ≤ 0.1A (230 nm, 3%, 1 cm): ≤ 0.2Note: Precipitation of SDS is not unusual. Warm
gently to redissolve.
77504 100 ml 500 ml 1 L
Sodium Dodecyl Sulfate (SDS), 20% SolutionULTRAPURE
Form: 20% (w/v) solution of sodium dodecyl sulfate (SDS) in high purity dH2O. Solution is 0.2 µm fil-tered and dispensed into durable, square Nalgene bottles.
A (280 nm, 3%, 1 cm): ≤ 0.1A (230 nm, 3%, 1 cm): ≤ 0.2Note: Precipitation of SDS is not unusual. Warm
gently to redissolve.
75832 100 ml 500 ml
Sodium Glutamate
See Glutamic Acid, Monosodium Salt, page 218
Sodium-N-Lauryl Sarcosine
See N-Lauryl Sarcosine, Sodium Salt, page 224
Sodium Lauryl Sulfate
See Sodium Dodecyl Sulfate, page 240
Sodium Phosphate, Dibasic Anhydrous
ACS Reagent GradeHygroscopicNa2HPO4
Formula Weight: 141.96Product specifications:Form: White crystalline powderAssay: ≥ 99.0%pH (5%, H2O, 25°C): 8.7 - 9.3Insoluble Matter: ≤ 0.01%Loss on Drying : ≤ 0.2%Chloride (Cl): ≤ 0.002%Sulfate (SO4): ≤ 0.005%Heavy Metals (Pb): ≤ 0.001%Iron (Fe): ≤ 0.002%Storage: Store ambient, protect from moisture
(product is hygroscopic).CAS #7558-79-4
21660 500 gm 2.5 kg
Sodium Phosphate, Dibasic, HeptahydrateULTRAPURE
ACS Reagent GradeEffloresces in warm dry airNa2HPO4
•7H2OFormula Weight: 268.07Product specifications:Form: White powderAssay: 98.0 - 102.0%pH (5%, H2O, 25°C): 8.7 - 9.3Insoluble Matter: ≤ 0.005%Chloride (Cl): ≤ 0.001%Nitrogen Compounds (N): ≤ 0.001%Sulfate (SO4): ≤ 0.005%Heavy Metals (Pb): ≤ 0.001%Arsenic (As): ≤ 5 ppmIron (Fe): ≤ 0.001%Note: While this product is a high quality, analytical
reagent grade buffer, as a good general labora-tory practice we recommend filtering solutions before use.
Storage: Store in tightly closed containers.CAS #7782-85-6
20232 500 gm 1 kg
Sodium Phosphate, Monobasic(Monosodium Phosphate)
CP GradeNaH2PO4
Formula Weight: 119.98Product specifications:Form: White, slightly hygroscopic crystalline powderAssay: 98.0 - 103.0% (NaH2PO4, dry basis)Identity: Passes testsArsenic (As): ≤ 3 ppmFluoride (F): ≤ 0.005%Insoluble Substances: ≤ 0.2%Lead (Pb): ≤ 4 ppmLoss on Drying: ≤ 2.0%Note: This product is a general laboratory reagent
suitable for most laboratory animal diets where trace element deficiencies are not required. One molar stock solutions of this product may be slightly hazy. Where use of a high quality research buffer is required, we recommend the use of our ACS Reagent Grade phosphate buffers.
Storage: Ambient, desiccate (product is slightly hygroscopic).
CAS #7558-80-7
20229 1 kg 5 kg
Biochemicals
241For bulk or alternate pack sizes, email us at [email protected].
Sodium Phosphate, Monobasic, MonohydrateULTRAPURE
ACS Reagent GradeSlightly HygroscopicNaH2PO4
•H2OFormula Weight: 137.99Product specifications:Form: White crystalline powderAssay: 98.0 - 102.0%pH (5%, H2O, 25°C): 4.1 - 4.5Insoluble Matter: ≤ 0.01%Chloride (Cl): ≤ 5 ppmSulfate (SO4): ≤ 0.003%Calcium (Ca): ≤ 0.005%Potassium (K): ≤ 0.01%Heavy Metals (Pb): ≤ 0.001%Iron (Fe): ≤ 0.001%CAS #10049-21-5
20233 500 gm 1 kg
Sodium Pyruvate
See Pyruvic Acid, Sodium Salt, page 237
Sodium Selenite
Na2SeO3
Formula Weight: 172.94Product specifications:Form: White to off-white powderStorage: Ambient; hygroscopic, store in tightly
closed containers in a cool, dry place. The com-pound is very readily reduced to red elemental selenium in the presence of small amounts of reducing agents, and should not be allowed to come into contact with organic material or even dust particles.
CAS #10102-18-8
21665 100 gm 500 gm
Sodium Succinate
See Succinic Acid, Disodium Salt, page 242
Soluble Starch
See Starch-Soluble, page 242
D-SorbitolULTRAPURE
USBioAnalyzedC6H14O6
Formula Weight: 182.17Product specifications:Form: White crystalline powderAssay (dry basis): 91.0 - 100.5%pH (10%, H2O): 3.5 - 7.0Identity: By IRIdentity (A, B): Passes testsMicrobial Limits: Passes testMoisture (KF): ≤ 1.5%Residue on Ignition: ≤ 0.1%Reducing Sugars: Passes testLimit of Nickel: Passes testStorage: In tightly closed containers, material is
hygroscopic.CAS #50-70-4
21710 500 gm 1 kg 5 kg
Spermidine, TrihydrochlorideULTRAPURE
C7H19N3•3HCl
Formula Weight: 254.63Product specifications:Form: White to pale pink crystalline powderAssay: ≥ 99%Identity: By IRApplications:� Causes a 2-fold increase in full length cDNA with
or without pyrophosphate or Actinomycin D using AMV Reverse Transcriptase.
� Used in cosmid cloning.� Often included in T4 Polynucleotide Kinase stocks.Storage: -20°CCAS #334-50-9
21760 1 gm 5 gm
Spermine, Tetrahydrochloride
C10H26N4•4HCl
Formula Weight: 348.18Product specifications:Form: White powderIdentity: By IRStorage: -20°CCAS #306-67-2
21755 5 gm
SSC, 20X SolutionULTRAPURE
Form: 20X SSC solution consisting of 0.3 M triso-dium citrate and 3.0 M sodium chloride in high purity dH2O, pH 7.0. Solution is 0.2 µm filtered and dispensed into durable, square Nalgene bottles. The 5 L size is supplied in an upright car-ton with a convenient dispensing spout.
pH (25°C, 20X): 7.0 ± 0.1Contaminant Testing: DNase (endo): Passes test DNase (exo): Passes test RNase: Passes testApplication: � Used in nucleic acid blotting and hybridization
techniques.
19629 500 ml 1 L 5 L
SSPE, 20X SolutionULTRAPURE
MB GradeForm: 20X SSPE solution consisting of 3 M NaCl, 200 mM sodium phosphate and 20 mM EDTA in
high purity dH2O, pH 7.4. Solution is 0.2 µm fil-tered and dispensed into durable, square Nalgene bottles. The 5 L size is supplied in an upright carton with a convenient dispensing spout.
pH (25°C, 20X): 7.4 ± 0.1Contaminant Testing: DNase (endo): Passes test DNase (exo): Passes test RNase: Passes test Protease: Passes testStorage: -20°C
75890 1 L 5 L
Biochemicals
242 888-362-2447 | 216-765-5000 | usb.affymetrix.com
Starch, HydrolyzedULTRAPURE
Source: PotatoProduct specifications:Form: White to off-white powderResidue on Ignition: ≤ 1%Loss on Drying: ≤ 20%Gel Strength: Passes testNote: Typically a 12-14% gel is recommended for
electrophoresis. Gel consistency is lot specific and depends on the percentage of amylose and amylopectin in starch, which varies naturally. Optimum concentration must be determined for each lot by the user.
CAS #9005-25-8
32823 100 gm 1 kg
Starch-SolubleULTRAPURE
ACS Reagent GradeSource: PotatoProduct specifications:Form: White powderSolubility: Passes testpH (2%, H2O, 25°C): 5.0 - 7.0Loss on Drying: ≤ 12%Sensitivity: Passes testResidue on Ignition: ≤ 0.4%CAS #9005-84-9
21695 1 kg
Stock Solutions
Ammonium Acetate (NH4OAc), 5 M Solution, page 199
Betaine, 5 M Solution, page 202Dithiothreitol (DTT) 0.1 M Solution, page 213EDTA, 0.5 M Solution, page 215Lithium Chloride (LiCl), 7.5 M Solution, page 225Magnesium Chloride (MgCl2), 1 M Solution,
page 226Potassium Chloride (KCl), 2 M Solution, page 236Sodium Acetate (NaOAc), 3 M Solution, pH 5.5,
page 239Sodium Chloride (NaCl), 5 M Solution, page 239Sodium Dodecyl Sulfate (SDS), 10% Solution,
page 240Sodium Dodecyl Sulfate (SDS), 20% Solution,
page 240Water, Nuclease-Free, page 250Water, RNase-Free, DEPC Treated, page 250
Stop SolutionULTRAPURE
Contents: 95% Formamide 20 mM EDTA 0.05% Bromophenol Blue 0.05% Xylene Cyanol FFApplication: � Used to stop and load sequencing reaction prod-
ucts onto denaturing acrylamide gels.Storage: -20°C
70724 1,200 µl
StreptavidinULTRAPURE
Source: E. coli recombinant proteinProduct specifications:Form: White, lyophilized powderActivity: ≥ 17 units/mgSolubility (0.1 M NaP, pH 7.4): ≥ 5 mg/mlSDS PAGE: Single bandUnit Definition: One unit is that amount of protein
required to bind one microgram of D-biotin.Applications:� A tetrameric protein containing four high affinity
binding sites for biotin.� Used with biotin in the purification of DNA-
binding proteins.Storage: 4°C, desiccated; -20°C, reconstituted.
11681 5 mg 10 mg
Streptavidin-Alkaline Phosphatase Conjugate
Storage: 4°C
11687 250 µl 1,500 µl
Streptomycin SulfateULTRAPURE
USBioAnalyzed(C21H39N7O12)2
•3H2SO4
Formula Weight: 1457.38Product specifications:Form: White to off-white powderActivity: 650 µg - 850 µg of C21H39N7O12/mg (as is)Identity (A, B): Passes testspH (200 mg streptomycin/ml): 4.5 - 7.0Loss on Drying: ≤ 5.0%Mode of Action: Inhibits protein synthesis by binding
to the S12 protein of the 30S ribosomal subunit.CAS #3810-74-0
21865 10 gm 100 gm 1 kg
Succinic Acid, Free Acid
HOOCCH2CH2COOHFormula Weight: 118.09Product specifications:Form: White crystalline powderAssay: ≥ 99.0%Identity: By IRMelting Point: 185 - 190°CMoisture (KF): ≤ 1.0%Iron (Fe): ≤ 5 ppmHeavy Metals: ≤ 10 ppmResidue on Ignition: ≤ 0.025%CAS #110-15-6
21900 500 gm 1 kg 5 kg
Succinic Acid, Disodium Salt, Hexahydrate(Sodium Succinate)
C4H4Na2O4•6H2O
Formula Weight: 270.14Product specifications:Form: White crystalline powderAssay: 97.0 - 102.0% (as is)Identity: By IRLoss on Drying: ≤ 40.0%Storage: Store in tightly closed containers.CAS #6106-21-4
21910 500 gm
Biochemicals
243For bulk or alternate pack sizes, email us at [email protected].
SucroseULTRAPURE
C12H22O11
Formula Weight: 342.30Product specifications:Form: White/clear crystalline powder[α]25
D : +66.3° to +66.8° (c=26 in H2O)A (260 nm, 1 M, 1 cm): ≤ 0.20Insoluble Matter: ≤ 0.005%Loss on Drying: ≤ 0.03%Residue on Ignition: ≤ 0.01%Titratable Acid: ≤ 0.0008 meq/gmInvert Sugar: ≤ 0.05%Chloride (Cl): ≤ 0.005%Sulfate and Sulfite (SO4): ≤ 0.005%Arsenic (As): ≤ 1 ppmBarium (Ba): ≤ 1 ppmCadmium (Cd): ≤ 1 ppmCalcium (Ca): ≤ 5 ppmCopper (Cu): ≤ 1 ppmIron (Fe): ≤ 1 ppmLead (Pb): ≤ 1 ppmMagnesium (Mg): ≤ 1 ppmManganese (Mn): ≤ 1 ppmZinc (Zn): ≤ 1 ppmContaminant Testing: DNase (endo): Passes test DNase (exo): Passes test RNase: Passes testCAS #57-50-1
21938 1 kg 5 kg
Sucrose
ACS Reagent GradeC12H22O11
Formula Weight: 342.30Product specifications:Form: White/clear crystalline powder[α]25
D : +66.3° to +66.8° (c=26, H2O)Insoluble Matter: ≤ 0.005%Loss on Drying: ≤ 0.03%Residue on Ignition: ≤ 0.01%Titratable Acid ≤ 0.0008 meq/gmChloride (Cl): ≤ 0.005%Sulfate and Sulfite (SO4): ≤ 0.005%Heavy Metals (Pb): ≤ 5 ppmIron (Fe): ≤ 5 ppmInvert Sugar: ≤ 0.05%CAS #57-50-1
21931 500 gm 2.5 kg
Sucrose, Crystalline
C12H22O11
Formula Weight: 342.30Product specifications:Form: White/clear crystalsIdentity: By IR[α]20
D : +65.9° to +67.1° (c=26 in H2O)Solubility (26%, H2O): Clear and colorlessCAS #57-50-1
21930 1 kg 5 lb 100 lb
Sulfanilic Acid
NH2C6H4SO3HFormula Weight: 173.19Product specifications:Form: White to pale cream colored powderAssay: 98.0 - 102.0% (as is)Identity: By IRCAS #121-57-3
21985 500 gm
TAE Buffer, 10X SolutionULTRAPURE
Form: 10X TAE solution consisting of 400 mM Tris and 0.01 M EDTA in high purity dH2O adjusted to pH 8.3. Solution is 0.2 µm filtered and dispensed into durable, square Nalgene bottles. The 5 L size is supplied in an upright carton with a convenient dispensing spout. Dilute to 1X with high purity dH2O for a solution consisting of 40 mM Tris Acetate, pH 8.3 and 1.0 mM EDTA.
pH (25°C, 10X): 8.3 ± 0.1Contaminant Testing: DNase (endo): Passes test DNase (exo): Passes test RNase: Passes test
75904 1 L 5 L
TAE Buffer, 50X SolutionULTRAPURE
Form: 50X TAE solution consisting of 2.0 M Tris and 0.05 M EDTA in high purity dH2O and adjusted to pH 8.3. Solution is 0.2 µm filtered and dispensed into durable, square Nalgene bottles. Dilute to 1X with high purity dH2O for a solution consisting of 40 mM Tris Acetate, pH 8.3 and 1.0 mM EDTA.
pH (25°C, 50X): 8.3 ± 0.1
74015 100 ml
Taurine(2-Amino Ethane Sulfonic Acid)
ULTRAPURE
MB GradeC2H7NO3SFormula Weight: 125.15Product specifications:Form: White crystalline powderAssay: ≥ 98.5%Identity: By IRSolubility (0.5 M, H2O): Clear and colorlesspH (5%, H2O): 4.0 - 7.0A (260 nm, 0.5 M, 1 cm): ≤ 0.2A (280 nm, 0.5 M, 1 cm): ≤ 0.2Residue on Ignition: ≤ 0.1%Heavy Metals: ≤ 10 ppmAmmonia (NH3): ≤ 0.02%Arsenic (As): ≤ 2 ppmChloride (Cl): ≤ 0.01%Sulfate (SO4): ≤ 0.01%Use Test: Functionally tested for electrophoresis.CAS #107-35-7
75824 1 kg
TBE Buffer, 5X SolutionULTRAPURE
Form: 5X TBE (Tris-Borate-EDTA) solution consisting of 0.445 M Tris, 0.445 M boric acid and 0.01M EDTA. Solution is 0.2 µm filtered and dispensed into durable, square Nalgene bottles. The 5 L size is supplied in an upright carton with a convenient dispensing spout. Dilute to 1X with high purity water for a solution consisting of 0.089 M Tris, 0.089 M boric acid and 0.002 M EDTA.
Testing: Meets or exceeds functional test criteria for high quality gel electrophoresis.
Application: � Primarily used as a buffer in gel electrophoresis
procedures.
75891 1 L 5 L
TBE Buffer, 10X Ready-Mixed PowderULTRAPURE
Form: Pre-weighed, pre-mixed powdered buffer packaged in 200 ml bottles for reconstitution to a 10X solution. Reconstitute with high purity dH2O to 200 ml. After reconstitution, the 10X solution consists of 0.89 M Tris, 0.89 M boric acid and
0.02 M EDTA. Each pack contains 6 x 200 ml bottles. Total volume is 1.2 liters of 10X buffer or 12 liters of 1X buffer.
Testing: Meets or exceeds functional test criteria for high quality gel electrophoresis.
Application: � Primarily used as a buffer in gel electrophoresis
procedures.Storage: Ambient. The reconstituted buffer is stable
for several days at 4°C if 0.2 µm filtered.
70454 6 x 200 ml
Biochemicals
244 888-362-2447 | 216-765-5000 | usb.affymetrix.com
TBS, 20X Solution, pH 7.4ULTRAPURE
MB GradeForm: 20X TBS (Tris Buffered Saline) solution con-
sisting of 500 mM Tris, 60 mM KCl and 2.8 M NaCl in high purity dH2O, pH 7.4. Solution is 0.2 µm filtered and dispensed into durable, square Nalgene bottles. The 5 L size is supplied in an upright carton with a convenient dispensing spout. Dilute to 1X with high purity water for a solution consisting of 25 mM Tris, pH 7.4, 3.0 mM KCl, and 140 mM NaCl.
pH (25°C, 20X): 7.4 ± 0.1Contaminant Testing: DNase (endo): Passes test DNase (exo): Passes test RNase: Passes test Protease: Passes testApplication: � Used in Western blotting protocols.
75892 100 ml 1 L 5 L
TBS with Tween (TBST), 20X Solution, pH 7.4ULTRAPURE
MB GradeForm: 20X TBS (Tris Buffered Saline) solution con-
sisting of 500 mM Tris, 60 mM KCl, 2.8 M NaCl, and 1.0% Tween 20 in high purity dH2O adjusted to pH 7.4. Solution is 0.2 µm filtered and is supplied in an upright carton with a convenient dispensing spout. Dilute to 1X with high purity water for a solution consisting of 25 mM Tris, pH 7.4, 3.0 mM KCl, 140 mM NaCl and 0.05% Tween 20.
pH (25°C, 20X): 7.4 ± 0.1Contaminant Testing: DNase (endo): Passes test DNase (exo): Passes test Protease: Passes testApplication: � Used in Western blotting protocols.
77500 1 L 5 L
TCEP-HCI, 0.5 M Solution, pH 6.6 ± 0.1(Tris [Carboxyethyl] Phosphine Hydrochloride)
Form: TCEP-HCI solution consisting of 0.5 M TCEP-HCI (Tris(2-Carboxyethyl)Phosphine Hydrochloride) in high purity dH2O.
pH (25°C): 6.6 ± 0.1Concentration (TCEP): 0.5 - 0.55 M
77531 5 ml 50 ml
TE Buffer, 1X SolutionULTRAPURE
MB GradeForm: 1X TE solution consisting of 10 mM Tris, pH
7.5, and 1 mM EDTA. Solution is 0.2 µm filtered and dispensed into vials and/or durable, square Nalgene bottles.
pH (25°C): 7.5 ± 0.1Contaminant Testing: DNase (endo): Passes test DNase (exo): Passes test RNase: Passes test Protease: Passes testApplication: � Use directly to resuspend and/or dilute purified
DNA or RNA.
75893 10 x 1 ml 100 ml 500 ml
TE Buffer, 1X Solution, pH 7.0
Form: 1X TE solution consisting of 10 mM Tris Ultrapure, pH 7.0, and 1 mM EDTA. Solution is 0.2 μm filtered and dispensed into vials and/or durable, square Nalgene bottles.
pH (25°C): 7.0 ± 0.1 Contaminant Testing: DNase (endo): Passes test DNase (exo): Passes test RNase: Passes test Protease: Passes testApplication:� Use directly to re-suspend and/or dilute purified
DNA or RNA.
75790 100 ml 500 ml
TE Buffer, 1X Solution, pH 7.0, Low EDTA
Form: 1X TE solution consisting of 10 mM Tris Ultrapure, pH 7.0, and 0.1 mM EDTA. Solution is 0.2 μm filtered and dispensed into vials and/or durable, square Nalgene bottles.
pH (25°C): 7.0 ± 0.1 Contaminant Testing: DNase (endo): Passes test DNase (exo): Passes test RNase: Passes test Protease: Passes testApplication:� Use directly to re-suspend and/or dilute purified
DNA or RNA.
75791 100 ml 500 ml
TE Buffer, 1X Solution, pH 8.0
MB GradeForm: 1X TE solution consisting of 10 mM Tris
Ultrapure, pH 8.0, and 1 mM EDTA. Solution is 0.2 μm filtered and dispensed into vials and/or durable, square Nalgene bottles.
pH (25°C): 8.0 ± 0.1 Contaminant Testing: DNase (endo): Passes test DNase (exo): Passes test RNase: Passes test Protease: Passes testApplication:� Use directly to re-suspend and/or dilute purified
DNA or RNA.
75792 100 ml 500 ml
TE Buffer, 1X Solution, pH 8.0, Low EDTA
MB GradeForm: 1X TE solution consisting of 10 mM Tris
Ultrapure, pH 8.0, and 0.1 mM EDTA. Solution is 0.2 μm filtered and dispensed into vials and/or durable, square Nalgene bottles.
pH (25°C): 8.0 ± 0.1 Contaminant Testing: DNase (endo): Passes test DNase (exo): Passes test RNase: Passes test Protease: Passes testApplication:� Use directly to re-suspend and/or dilute purified
DNA or RNA.
75793 100 ml 500 ml
TE Buffer, 50X SolutionULTRAPURE
Form: 50X TE solution consisting of 500 mM Tris, 50 mM EDTA in high purity dH2O, adjusted to pH 7.5. Solution is 0.2 µm filtered and dispensed into durable, square Nalgene bottles. Dilute to 1X with high purity dH2O for a solution consisting of 10 mM Tris, pH 7.5, and 1 mM EDTA.
pH (25°C, 50X): 7.5 ± 0.1Contaminant Testing: DNase (endo): Passes test DNase (exo): Passes test RNase: Passes test Protease: Passes test
75834 100 ml 500 ml
Biochemicals
245For bulk or alternate pack sizes, email us at [email protected].
TEMED
See Tetramethylethylenediamine, page 245
Terrific Broth, Ready-Made Powder
Form: Off-white free flowing powder.pH (5.08%, H2O): 7.2 ± 0.2Contents: Casein Peptone: 12 gm/L Yeast Extract: 24 gm/L Potassium Phosphate Dibasic: 12.5 gm/L Potassium Phosphate Monobasic: 2.3 gm/LApplication: � Provides greater yields of plasmid DNA and
recombinant proteins than LB media by allowing for greater cell density.
To use: Add 50.8 gm powder, 4 ml glycerol, and 996 ml water to make 1 liter medium. Dissolve and autoclave.
75856 250 gm 1 kg
TES(N-Tris[Hydroxymethyl]methyl-2-Aminoethanesulfonic Acid)
ULTRAPURE
C6H15NO6SFormula Wt.: 229.25Product specifications:Form: White/clear crystalline powderAssay: ≥ 98.5% (dry basis)Identity A: KBr pellet matches referenceIdentity B: Passes testpKa (20°C): 7.50 ± 0.15A (250 nm, 1 M, 1 cm): ≤ 0.2Loss on Drying: ≤ 1.0%Iron (Fe): ≤ 30 ppmLead (Pb): ≤ 10 ppmCAS #7365-44-8
22086 100 gm 500 gm 1 kg
Tetracycline, HydrochlorideULTRAPURE
USBioAnalyzedC22H24N2O8
•HClFormula Weight: 480.90Product specifications:Form: Yellow crystalline powderActivity: ≥ 900 µg of C22H24N2O8
•HCI/mg (as is)Identity (A, B, C, D, E, F): Passes testsCrystallinity: Passes testpH (10 mg/ml): 1.8 - 2.8[α]25
D : -240° to -255° (c=0.5, 0.1N HCI, dry basis)Loss on Drying (60°C, 3 hours, vacuum): ≤ 2.0%Limit of 4-Epianhydrotetracycline: ≤ 2.0%Heavy Metals (Pb): ≤ 0.005%Storage: Ambient in tightly closed, light-resistant
containers.CAS #64-75-5
22105 25 gm 100 gm 1 kg
Tetramethylammonium Chloride(TMAC)
ULTRAPURE
C4H12ClNFormula Weight: 109.60Product specifications:Form: White crystalline powderAssay: ≥ 98% (dry basis)Identity: By IRpH (10%, H2O): ≥ 4.5Loss on Drying: ≤ 1.5%Solubility (10%, H2O): Clear and colorlessApplications:� Used in hybridization solutions of mixed oligo-
nucleotides. In solutions with TMAC, the Tm (melt-ing temperature) of a hybrid is dependent on its length, not its base composition.
� Used in probing PCR products for the analysis of polymorphisms with allele-specific oligonucle-otides.
Storage: Store at room temperature. This material is extremely hygroscopic.
CAS #75-57-0
22124 1 kg
3,3',5,5'-Tetramethylbenzidine(TMB)
C16H20N2
Formula Weight: 240.35Product specifications:Form: Off-white powderIdentity: By IRMelting Point: 168.3 - 169.9°CCAS #54827-17-7
22126 1 gm 5 gm
N,N,N',N'-Tetramethylethylene diamine(TEMED)
ULTRAPURE
MB Grade(CH3)2NCH2CH2N(CH3)2
Formula Weight: 116.20Product specifications:Form: Colorless to pale yellow liquidAssay: ≥ 98%A (290 nm, 1%, 1 cm): ≤ 0.1Density (20°C): 0.77 ± 0.01Refractive Index (20°C): 1.419 ± 0.001Testing: Meets or exceeds functional test criteria for
high quality gel electrophoresis.Application: � Catalyzes the formation of free radicals by ammo-
nium persulfate and accelerates the polymeriza-tion of acrylamide and bisacrylamide.
CAS #110-18-9.
76320 100 ml 100 gm 500 gm
Tetrazolium Red
See 2,3,5-Triphenyl Tetrazolium Chloride, page 246
Thiamine, Hydrochloride(Vitamin B1 ; Aneurine)
USBioAnalyzedC12H17CIN4OS•HClFormula Weight: 337.27Product specifications:Form: White crystalline powderAssay: 98.0 - 102.0% (dry basis)Identity (A, B): Passes testsChromatographic Purity: Passes testpH (1%, H2O): 2.7 - 3.4Spectral Ratios (400 nm, 10%, 1 cm): ≤ 0.025Moisture (KF): ≤ 5.0%Residue on Ignition: ≤ 0.2%Nitrate: Passes testResidual Solvents: Meets the requirementsCAS #67-03-8
22170 100 gm 500 gm 1 kg
Thiamine Pyrophosphate Chloride
See Cocarboxylase, page 207
Thiazolyl Blue
See MTT, page 228
Biochemicals
246 888-362-2447 | 216-765-5000 | usb.affymetrix.com
Thimerosal
USBioAnalyzedC9H9HgNaO2SFormula Weight: 404.81Product specifications:Form: White powderAssay: 97.0 - 101.0% (dry basis)Identity (A, B): Passes testsLoss on Drying: ≤ 0.5%Ether-Soluble Substances: Passes testMercury Ions: ≤ 0.70%Readily Carbonizable Substances: Passes testStorage: Store in light-resistant containers.CAS #54-64-8
22215 25 gm 100 gm
Thioredoxin, Human, Recombinant(r-hTRX)
Formula Weight: 11,734 determined by amino acid sequence
Product specifications:Form: Lyophilized powder from phosphate bufferSpecific Activity: ≥ 3 units/mg proteinUnit Definition: TRX activity is assayed by measuring
the change in absorbance at 650 nm at 25°C using 0.13 µM bovine insulin containing 0.33 mM DTT (pH 6.5).
Storage: -20°C, protect from moisture.
26004 100 µg
L-Threonine
CH3CH(OH)CH(NH2)COOHFormula Weight: 119.12Product specifications:Form: White crystalline powderAssay: ≥ 98.5%Identity: By IRpH (5%, H2O): 5.0 - 6.5[α]25
D : -26.7° to -29.1° (c=6 in H2O)Loss on Drying: ≤ 0.2%Residue on Ignition: ≤ 0.4%Heavy Metals (Pb): ≤ 15 ppmChloride (Cl): ≤ 0.05%Iron (Fe): ≤ 30 ppmSulfate (SO4): ≤ 0.03%Storage: Store in light-resistant containers.CAS #72-19-5
22290 100 gm 1 kg
Thrombin Human
Source: Human1,000 NIH units/vialProduct specifications:Form: Lyophilized powderActivity: ≥ 1,000 NIH units/vialSpecific Activity: ≥ 2,000 NIH units/mg proteinReconstitution: Reconstitute with 1.0 ml ddH2OBuffer: After reconstitution the solution contains: 5 mM Sodium Citrate 0.01% PEG 0.02 M NaCl pH 6.5All source material tested and found non-reactive
to HTLV-III and HBsAg using FDA approved assay methods.
Storage: -20°C
22292 1,000 units
Thrombin
(E.C.3.4.21.5)Source: Bovine BloodProduct specifications:Form: Lyophilized powderActivity: ≥ 40 units/mg powderMoisture: ≤ 5.0%Note: The activity is determined by comparison
to the activity of Standard Thrombin from the Bureau of Biological Standards.
Storage: 4°C, in tightly closed containers.CAS #9002-04-4
22293 25,000 units 100,000 units
TMAC
See Tetramethylammonium Chloride, page 245
TMB
See 3,3',5,5'-Tetramethylbenzidine, page 245
Toxillic Acid
See Maleic Acid, page 226
α,α-Trehalose, Dihydrate(α-D-Glucopyranosyl-α-D-Glucopyranoside)
C12H22O11•2H2O
Formula Weight: 378.33Product specifications:Form: White crystalline powderMoisture (KF): 9.3 - 9.7%[α]20
D : +179.0° ± 1.5° (c=7, H2O)Arsenic (As): ≤ 0.5 ppmIron (Fe): ≤ 5 ppmHeavy Metals (Cu): ≤ 10 ppmCAS #6138-23-4
22515 100 gm
Tricine[N-Tris (Hydroxymethyl)methylglycine]
ULTRAPURE
(CH2OH)3CNHCH2COOHFormula Weight: 179.17Product specifications:Form: White/clear crystalline powderAssay: ≥ 98.5% (dry basis)Identity: By IRpKa (20°C): 8.15 ± 0.15A (250 nm, 5%, 1 cm): ≤ 0.1Loss on Drying: ≤ 0.5%Iron (Fe): ≤ 30 ppmLead (Pb): ≤ 10 ppmCAS #5704-04-1
22561 100 gm 500 gm 1 kg
Triethanolamine, Hydrochloride
N(CH2CH2OH)3•HCl
Formula Weight: 185.65Product specifications:Form: White/clear crystalline powderAssay: ≥ 98.5% (dry basis)Identity: By IRpKa (20°C): 7.85 ± 0.15Melting Point: 178 - 182°CLoss on Drying: ≤ 1%Iron (Fe): ≤ 30 ppmHeavy Metals (Pb): ≤ 10 ppmCAS #637-39-8
22565 250 gm 1 kg
1,3,7-Trimethylxanthine
See Caffeine, page 204
2,6,8-Trioxypurine
See Uric Acid, page 249
2,3,5-Triphenyl Tetrazolium Chloride(TTC; Tetrazolium Red)
C19H15ClN4
Formula Weight: 334.80Product specifications:Form: Cream to beige colored powderIdentity: By IRSolubility (1%, H2O): Clear solutionMoisture: ≤ 5% (By KF)Sensitivity for Reducing Sugars: Passes testCAS #298-96-4
22631 10 gm 25 gm 50 gm 100 gm 500 gm 1 kg
Biochemicals
247For bulk or alternate pack sizes, email us at [email protected].
Triphosphopyridine Nucleotide Dinucleotide
See Nicotinamide Adenine Dinucleotide Reduced, page 230
2,4,6-Tripyridyl-s-Triazine (TPTZ)
C18H12N6
Formula Weight: 312.33CAS #3682-35-7
22660 5 gm 25 gm 100 gm
Tris(Tris[hydroxymethyl]aminomethane)
ULTRAPURE
MB GradeNH2C(CH2OH)3
Formula Weight: 121.14Product specifications:Form: White crystalline powderAssay: 99.8 - 100.1% (dry basis)Identity: By IRpKa (20°C): 8.30 ± 0.15A (250 nm, 1 M, 1 cm): ≤ 0.1A (290 nm, 40%, 1 cm): ≤ 0.2Solubility (1 M, 500 ml): Clear and colorlesspH (5%, H2O): 10.0 - 11.5Melting Point: 168 - 172°CLoss on Drying: ≤ 1%Moisture (KF): ≤ 1%Residue on Ignition: ≤ 0.1%Insoluble Matter: ≤ 0.005%Arsenic (As): ≤ 1 ppmBarium (Ba): ≤ 1 ppmCadmium (Cd): ≤ 1 ppmCalcium (Ca): ≤ 1 ppmCopper (Cu): ≤ 1 ppmIron (Fe): ≤ 1 ppmLead (Pb): ≤ 1 ppmMagnesium (Mg): ≤ 1 ppmManganese (Mn): ≤ 1 ppmZinc (Zn): ≤ 1 ppmDNase (endo): Passes testDNase (exo): Passes testRNase: Passes testProtease: Passes testCAS #77-86-1
75825 500 gm 1 kg 5 kg 10 kg
Tris[Tris(hydroxymethyl)aminomethane]
High Purity GradeNH2C(CH2OH)3
Formula Weight: 121.14Product specifications:Form: White/clear crystalline powderAssay: 99.0 - 101.0% (dry basis)Identity: By IRpKa (20°C): 8.30 ± 0.15A (250 nm, 1 M, 1 cm): ≤ 0.1A (290 nm, 40%, 1 cm): ≤ 0.2pH (5%, H2O): 10.0 - 11.5Melting Point: 168 - 178°CLoss on Drying: ≤ 1%Residue on Ignition: ≤ 0.1%Insoluble Matter: ≤ 0.005%Iron (Fe): ≤ 5 ppmHeavy Metals (Pb): ≤ 5 ppmCAS #77-86-1
22674 500 gm 1 kg 5 kg
Tris[Tris(hydroxymethyl)aminomethane]
Reagent GradeNH2C(CH2OH)3
Formula Weight: 121.14Product specifications:Form: White to pale yellow crystalline powderAssay: ≥ 98.0% (dry basis)Identity: By IRpH (5%, H2O): 10.0 - 11.5Melting Point: ≥ 160°CLoss on Drying: ≤ 1.5%CAS #77-86-1
22675 500 gm 1 kg
Tris-Glycine (TG) Buffer, 10X SolutionULTRAPURE
MB GradeForm: 10X Tris-Glycine (TG) Buffer consisting of 0.25 M Ultrapure Tris, pH 8.3, and 1.92 M gly-
cine. Solution is 0.2 µm filtered and dispensed into durable, square Nalgene bottles. The 5 L size is supplied in an upright carton with a convenient dispensing spout. Dilute to 1X with high purity water and methanol for a solution consisting of 25 mM Tris, pH 8.3, 192 mM glycine, 20% methanol.
Contaminant Testing: Protease: Passes test.Application: � For Western blotting and gel electrophoresis.
75894 1 L 5 L
Tris-Glycine-SDS (TGS) Buffer, 10X SolutionULTRAPURE
MB GradeForm: 10X Tris-Glycine-SDS (TGS) Buffer, consisting
of 0.25 M Ultrapure Tris, pH 8.6, 1.92 M glycine, and 1.0% SDS. Solution is 0.2 µm filtered and dispensed into durable, square Nalgene bottles. The 5 L size is supplied in an upright carton with a convenient dispensing spout. Dilute to 1X with high purity dH2O for a solution consisting of 25 mM Tris, pH 8.6, 192 mM glycine and 0.1% SDS.
Contaminant Testing: Protease: Passes testApplication: � TGS is the most commonly used buffer for SDS-
PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) of proteins.
75895 1 L 5 L
Tris, Hydrochloride(Tris[hydroxymethyl]aminomethane Hydrochloride)
ULTRAPURE
MB GradeC4H11NO3 HClFormula Weight: 157.60Product specifications:Form: White crystalline powderAssay: 99.0 - 101.0% (as is)Identity: By IRpKa (20°C): 8.30 ± 0.15A (250 nm, 1 M, 1 cm): ≤ 0.20A (290 nm, 40%, 1 cm): ≤ 0.20pH (5%, H2O): 4.7 ± 0.5Melting Point: 148 - 157°CLoss on Drying: ≤ 1.0%Residue on Ignition: ≤ 0.1%Insoluble Matter: ≤ 0.005%Iron (Fe): ≤ 5 ppmHeavy Metals (Pb): ≤ 5 ppmDNase (endo): Passes testDNase (exo): Passes testRNase: Passes testProtease: Passes testStorage: Store in tightly closed containers.CAS #1185-53-1
22676 500 gm 1 kg 5 kg
Biochemicals
248 888-362-2447 | 216-765-5000 | usb.affymetrix.com
Tris/HCl, 1 M Solution, pH 7.0ULTRAPURE
MB GradeForm: 1 M solution of Tris in high purity dH2O,
adjusted to pH 7.0. Solution is 0.2 µm filtered and dispensed into durable, square Nalgene bottles.
pH (25°C): Passes testContaminant Testing: DNase (endo): Passes test DNase (exo): Passes test RNase: Passes test Protease: Passes testCAS #1185-53-1
22637 100 ml 500 ml 1 L
Tris/HCl, 1 M Solution, pH 7.5ULTRAPURE
MB GradeForm: 1 M solution of Tris in high purity dH2O,
adjusted to pH 7.5. Solution is 0.2 µm filtered and dispensed into durable, square Nalgene bottles.
pH (25°C): 7.5 ± 0.1Contaminant Testing: DNase (endo): Passes test DNase (exo): Passes test RNase: Passes test Protease: Passes testCAS #1185-53-1
22639 100 ml 500 ml 1 L
Tris/HCl, 1 M Solution, pH 8.0ULTRAPURE
MB GradeForm: 1 M solution of Tris, Ultrapure in high purity
dH2O, adjusted to pH 8.0. Solution is 0.2 µm fil-tered and dispensed into durable, square Nalgene bottles.
pH (25°C): Passes testContaminant Testing: DNase (endo): Passes test DNase (exo): Passes test RNase: Passes test Protease: Passes test
22638 100 ml 500 ml 1 L
r-hTRX
See Thioredoxin, Human, Recombinant, page 246
TrypsinULTRAPURE
(E.C.3.4.21.4)USBioAnalyzedSource: Bovine PancreasProduct specifications:Form: White to pale yellow powderActivity: ≥ 2,500 USP units/mg powderContaminating Activity: Chymotrypsin: ≤ 50 USP units/mg powderLoss on Drying: ≤ 5.0%Solubility: Passes testResidue on Ignition: ≤ 2.5%Microbial Limits Test per USP: Passes testStorage: -20°C, protect from moisture.CAS #9002-07-7
22720 1 gm 5 gm 10 gm
Trypsin (TPCK Treated)(E.C.3.4.21.4)Source: Bovine PancreasProduct specifications:Form: Lyophilized powder, treated with TPCK to
inhibit chymotryptic activityActivity: ≥ 180 units/mg proteinUnit Definition: One unit hydrolyzes 1 µmol of
TAME/minute at 25°C, pH 8.2, in the presence of 0.01 M Ca2+.
Storage: 4°CCAS #9002-07-7
22725 100 mg 250 mg
Trypsin 1:250
(E.C.3.4.21.4)Source: Porcine PancreasProduct specifications:Form: Powder stabilized with lactose.Activity: ≥ 250,000 USP units/gmChymotrypsin: ≥ 75,000 USP units/gmUnit Definition: One unit causes a change in absor-
bance of 0.003/minute with BAEE as a substrate at 25°C, pH 7.6.
Storage: 4°CCAS #9002-07-7
22710 100 gm 1 kg
L-Tryptophan(L-2-Amino-3-Indole Propionic Acid)
USBioAnalyzedC11H12N2O2
Formula Weight: 204.23Product specifications:Form: White to off-white crystalline powderAssay: 98.5 - 101.5% C11H12N2O2 (dry basis)Identity: By IRpH (1%, H2O): 5.5 - 7.0Chromatography (TLC): Homogeneous[α]25
D : -29.4° to -32.8° (c=1 in H2O)Loss on Drying: ≤ 0.3%Residue on Ignition: ≤ 0.1%Heavy Metals (Pb): ≤ 0.0015%Chloride (Cl): ≤ 0.05%Iron (Fe): ≤ 0.003%Sulfate (SO4): ≤ 0.03%Residual Solvents: Meets the requirementsCAS #73-22-3
22765 100 gm 1 kg
Tyrosinase(Polyphenol Oxidase)
(E.C.1.14.18.1)Source: MushroomProduct specifications:Form: Lyophilized powderActivity: ≥ 500 units/mg (dry basis)Storage: -20°C
22885 100,000 units 500,000 units
L-Tyrosine
USBioAnalyzedHO(C6H4)CH2CH(NH2)COOHFormula Weight: 181.19Product specifications:Form: White to off-white crystalline powderAssay: 98.5 - 101.5% (dry basis)Identity: By IR[α]25
D : -9.8° to -11.2° (dry basis, c=5 in 1N HCl)Loss on Drying (105°C, 3 hours): ≤ 0.3%Residue on Ignition: ≤ 0.4%Chloride (Cl): ≤ 0.04%Iron (Fe): ≤ 30 ppmSulfate (SO4): ≤ 0.04%Heavy Metals (Pb): ≤ 15 ppmChromotographic Purity: Passes testResidual Solvents: Meets the requirementsCAS #60-18-4
22910 100 gm 1 kg 5 kg
Biochemicals
249For bulk or alternate pack sizes, email us at [email protected].
Uracil
C4H4N2O2
Formula Weight: 112.09Product specifications:Form: White to off-white crystalline powderAssay (Total UV, pH 7.0): ≥ 98.0% (dry basis)Identity: By IRSpectral Ratios (pH 7.0): A280/A260: 0.18 ± 0.02 A250/A260: 0.84 ± 0.03Loss on Drying (105°C, 2 hours): ≤ 5.0%Residue on Ignition: ≤ 0.1%Reference constants:λmax (pH 7, NRC): 260 nmεmax (pH 7, NRC): 8.2 x 103
ε260 (pH 7, DBR): 8.2 x 103
CAS #66-22-8
23020 100 gm 1 kg
UreaULTRAPURE
MB GradeNH2CONH2
Formula Weight: 60.06Product specifications:Form: White crystalline powderAssay: ≥ 99.5%Identity: By IRSpectral Ratios (260 nm, 6 M, 1 cm): ≤ 0.055Spectral Ratios (280 nm, 6 M, 1 cm): ≤ 0.044Melting Point: 132 - 135°CInsoluble Matter ≤ 0.01%Residue on Ignition: ≤ 0.01%Conductivity: ≤ 15 µmhos/cmCyanate: None detectedChloride (Cl): ≤ 0.0005%Sulfate (SO4): ≤ 0.001%Copper (Cu): ≤ 0.5 ppmIron (Fe): ≤ 0.5 ppmLead (Pb): ≤ 0.5 ppmDNase (endo): Passes testRNase: Passes testProtease: Passes testApplications:� Chaotropic agent used in denaturing polyacryla-
mide gel electrophoresis.� When 8 M urea is not sufficiently denaturing,
40% formamide may be included.CAS #57-13-6
75826 1 kg 5 kg
Urea
High Purity GradeNH2CONH2
Formula Weight: 60.06Product specifications:Form: White crystalline powderAssay: 99.0 - 100.5%Identity: By IRLoss on Drying: ≤ 1.0%Melting Point: 132 - 135°CInsoluble Matter: ≤ 0.01%Residue on Ignition: ≤ 0.01%Chloride (Cl): ≤ 5 ppmSulfate (SO4): ≤ 0.001%Heavy Metals (Pb): ≤ 0.001%Iron (Fe): ≤ 0.001%CAS #57-13-6
23036 1 kg 5 kg
Uric Acid(2,6,8-Trioxypurine)
C5H4N4O3
Formula Weight: 168.11Product specifications:Form: White to off-white powderAssay: ≥ 98.5%Identity: By IRSolubility: Clear solutionLoss on Drying: ≤ 0.2%Residue on Ignition: ≤ 0.2%Heavy Metals: ≤ 20 ppmCAS #69-93-2
23075 100 gm 500 gm 1 kg
Uridine
C9H12N2O6
Formula Weight: 244.20Product specifications:Form: White to slightly off-white powderAssay (UV): 100% ± 2%Identity: By IRλmax (pH 7.0): 262 nm ± 1 nmεmax (pH 7.0): [10.1 ± 2%] x 103
Spectral Ratios (pH 7.0): A250/A260: 0.74 ± 0.03 A280/A260: 0.36 ± 0.02Loss on Drying: ≤ 1.0%Heavy Metals (Pb): ≤ 10 ppmReference constants:λmax (pH 7, NRC): 262 nmεmax (pH 7, NRC): 10.1 x 103
ε260 (pH 7, DBR): 10.0 x 103
CAS #58-96-8
23130 1 gm 10 gm 100 gm
Uridine-5'-Triphosphate, Trisodium Salt(UTP)
C9H12N2O15P3Na3•2H2O
Formula Weight: 586.12Product specifications:Form: White crystalline powderAssay (Total UV): 100% ± 3% (dry basis)Assay (HPLC): ≥ 90%λmax (pH 7.0): 262 nm ± 1 nmεmax (pH 7.0): [10.0 ± 0.3] x 103
Spectral Ratios (pH 7.0): A250/A260: 0.60 - 0.90 A280/A260: 0.30 - 0.45Moisture: ≤ 15.0%Reference constants:λmax (pH 7, NRC): 262 nmεmax (pH 7, NRC): 10.0 x 103
ε260 (pH 7, DBR): 9.9 x 103
Storage: -20°CCAS #19817-92-6
23160 100 mg 1 gm
UTP
See Uridine-5'-Triphosphate, page 249
Biochemicals
250 888-362-2447 | 216-765-5000 | usb.affymetrix.com
Vancomycin, Hydrochloride
USBioAnalyzedC66H75Cl2N9O24
•HClFormula Weight: 1485.71Product specifications:Form: Tan to brown powderActivity: ≥ 900 µg of vancomycin/mg (dry basis) ≥ 855 µg of vancomycin/mg (as is)Identity: By IRChromatographic Purity: Vancomycin B: ≥ 80.0% Other Peaks: ≤ 9%pH (50 mg/ml, H2O): 2.5 - 4.5Moisture (KF): ≤ 5.0%Storage: 4°C, in tightly closed containers.CAS #1404-93-9
23261 100 mg 250 mg
Vitafree Casein
See Casein Vitafree, page 205
Vitamin B1
See Thiamine HCl, page 245
Vitamin B2
See Riboflavin, page 238
Vitamin B5
See D-Calcium Pantothenate, page 204
Vitamin B12
(Cyanocobalamin)
USBioAnalyzedC63H88CoN14O14PFormula Weight: 1355.37Product specifications:Form: Dark red crystalline powderAssay: 96.0 - 100.5% (dry basis)Identification A: Passes testIdentification B: Passes testIdentification C: Passes testLoss on Drying: ≤ 12.0%Psuedocyanocobalamin: Passes testStorage: In tightly closed, light-resistant containers.CAS #68-19-9
23360 1 gm
Vitamin C
See L-Ascorbic Acid, page 201
Vitamin Free Casein
See Casein-Vitafree, page 205
Vitamin G
See Riboflavin, page 238
Vitamin K1
(Phytonadione; 2-methyl-3-Phytyl-3-1; 4-Napthoquinone)
USBioAnalyzedC31H46O2
Formula Weight: 450.70Product specifications:Form: Clear yellow to amber viscous liquidAssay: 97.0 - 103.0%Identity (A, B): Passes testsRefractive Index (25°C): 1.523 to 1.526Reaction: Neutral to litmusLimit of Menadione: Passes testZ-isomer content: ≤ 21.0%CAS #84-80-0
23445 250 mg 1 gm 5 gm 10 gm
Water, Nuclease-FreeULTRAPURE
MB GradeForm: High purity dH2O tested for contaminating
nuclease activity. Product is 0.2 µm filtered and dispensed into vials and/or durable, square Nalgene bottles. The 5 L size is supplied in an upright carton with a convenient dispensing spout.
Contaminant Testing: RNase: Passes test DNase (endo): Passes test DNase (exo): Passes test Protease: Passes testAssays: Functional Assay: Functionally tested for PCR
according to the standard Affymetrix PCR protocol.
Enzyme Inhibition Assay: No inhibition by nuclease-free H2O on digestion of lambda DNA by Hind III.
71786 10 x 1 ml 100 ml 500 ml 1 L 5 L
Water, RNase-Free, DEPC TreatedULTRAPURE
MB GradeForm: High purity dH2O treated overnight with 0.1%
Diethylpyrocarbonate. Product is 0.2 µm filtered and dispensed into vials and/or durable, square Nalgene bottles, then autoclaved.
Contaminant Testing: RNase: Passes test DNase (endo): Passes test DNase (exo): Passes test Protease: Passes testAssays: Functional Assay: Functionally tested for PCR
according to the standard Affymetrix PCR protocol.
Enzyme Inhibition Assay: No inhibition by RNase-free H2O on digestion of lambda DNA by Hind III.
70783 25 ml 10 x 1 ml 100 ml 500 ml 1 L
Wright’s Stain
Prepared for use in blood stain work.CAS #68988-92-1
23457 5 gm 10 gm 25 gm 100 gm
Xanthine Monosodium Salt, Monohydrate
C5H3N4O2Na•H2OFormula Weight: 192.11Product specifications:Form: White to off-white crystalline powderAssay (Total UV): ≥ 96.0% (dry basis)Identity: By IRMoisture (KF): ≤ 12.0%Spectral Ratios (pH 10.0): A250/A260: 1.29 ± 0.06 A280/A260: 1.71 ± 0.05Reference constants:λmax (pH 10, NRC): 277 nmεmax (pH 10, NRC): 9.3 x 103
ε260 (pH 10, DBR): 5.2 x 103
CAS #1196-43-6
23465 5 gm 10 gm 25 gm 100 gm
Biochemicals
251For bulk or alternate pack sizes, email us at [email protected].
X-α-GAL
See 5-Bromo-4-Chloro-3-Indolyl-α-D-Galactopyranoside, page 203
X-Gal
See 5-Bromo-4-Chloro-3-Indolyl-β-D-Galactopyranoside, page 203
Xylene Cyanol FFULTRAPURE
(C.I.43535)C25H27N2NaO7S2
Formula Weight: 554.61Product specifications:Form: Dark burgundy metallic powderIdentity: By IRεmax (615 nm, H2O, 1 cm): ≥ 60 x 103
Loss on Drying: ≤ 10.0%λmax (H2O solution): 615 nm ± 2 nmApplication:� Useful as a tracking dye during the separation of
nucleic acids (in particular, in DNA sequencing electrophoresis).
Sequencing Stop Solution: 95% Formamide 20 mM EDTA 0.05% Bromophenol Blue 0.05% Xylene Cyanol FF Store solution at -20°C.10X Agarose Gel Loading Solution: 50% Glycerol 60 mM EDTA 0.05% Bromophenol Blue 0.05% Xylene Cyanol 1.0% SDSStorage: Store solution at room temperature.CAS #4463-44-9
23513 25 gm
D-(+)-Xylose
C5H10O5
Formula Weight: 150.13Product specifications:Form: White crystalline powderAssay: ≥ 98.5%[α]20
D : +18.9° to +19.7° (c=4 in H2O)Loss on Drying: ≤ 1.0%Residue on Ignition: ≤ 0.05%Heavy Metals (Pb): ≤ 5 ppmCAS #58-86-6
23520 500 gm 1 kg
Yeast, Bakers
Source: Baker’s YeastStorage: 4°C
23540 1 lb 5 lb 25 lb
Yeast, Brewers
CAS #68876-77-7
23546 5 lb 25 lb 100 lb
Yeast Extract Powder(Peptones)
ULTRAPURE
Source: Baker’s YeastProduct specifications:Form: Off-white powderProtein (TN x 6.25): 57-72%Moisture: ≤ 6% at time of packagingSalmonella: NegativeStandard Plate Count: ≤ 15,000/gmThis product is hygroscopic.Storage: Ambient, in tightly closed containers.CAS #8013-01-2
23547 1 kg
Yeast Extract Powder, Low Dust (Peptones)
Source: Baker’s YeastProduct specifications:Form: Tan powder, low dustProtein (TN x 6.25): 57 - 72%Moisture: ≤ 6% at time of packagingSalmonella: NegativeStandard Plate Count: ≤ 15,000/gmThis product is hygroscopic.Storage: Ambient, in tightly closed container.CAS #8013-01-2
23545 1 kg 5 kg
Yeast Hydrolysate, Enzymatic(Peptones)
ULTRAPURE
Product specifications:Form: Light tan powderIdentity: By IRSolubility (5%, H2O): Passes testpH (5% H2O): 5.4 - 6.2CAS #100684-36-4
23550 5 lb 25 lb 50 lb 100 lb
Yeast-Torula
23551 5 lb 25 lb 100 lb
2X YT Broth
Form: Off-white, free-flowing powderpH (3.1%, H2O): 7.0 ± 0.2Contents: Casein Peptone: 16 gm/L Yeast Extract: 10 gm/L Sodium Chloride: 5 gm/LTo Use: Dissolve 31 gm powder per liter of purified
water.
75864 1 kg
Code Product Size Page Code Product Size Page
Product Code Index | Numerical
253For bulk or alternate pack sizes, email us at [email protected].
10062 S-AcetylCoenzymeA,LithiumSalt,Ultrapure 10mg 188 25mg 100mg 500mg
10077 5-Bromo-4-Chloro-3-Indolyl-b-D-Galactopyranoside, 100mg 203 Ultrapure 250mg 1gm 2gm 5gm
10120 N-Acetyl-L-Cysteine 100gm 188 500gm 1kg
10132 Agarose-Hi-Res,Separation≤1000bp,Ultrapure 25gm 195 100gm 500gm
10430 Adenine 100gm 188 1kg
10450 AdenineSulfate,Dihydrate 100gm 188 1kg
10460 Adenosine 100gm 189
10490 Adenosine-5’-Diphosphate,DisodiumSalt,Dihydrate 5gm 189 10gm 25gm
10520 Adenosine-5’-Monophosphate,FreeAcid 10gm 189 25gm 100gm
10521 Adenosine-5’-Monophosphate,FreeAcid,Monohydrate 25gm 189 100gm 1kg
10585 Adenosine-5’-Triphosphate,DisodiumSalt 5gm 189 10gm 25gm 100gm 500gm
10601 S-Adenosyl-L-MethionineSulfatep-Toluenesulfonate 25mg 190 100mg
10620 Adenosine-5’-Monophosphate,DisodiumSalt 25gm 189 100gm
10635 Adonitol 25gm 190 100gm
10654 Agar 1lb 190 5lb 25lb
10665 b-Alanine 200gm 198 1kg 5kg
10848 Albumin,Bovine,Acetylated,Ultrapure 25mg 198 125mg
10852 Albumin,Bovine,CohnFractionV,pH5.2 50gm 198 100gm 500gm
10856 Albumin,Bovine,Crystallized,pH5.2 5gm 198 10gm 100gm
10857 Albumin,Bovine,pH7 50gm 198 100gm 500gm 1kg
10865 Albumin,Egg 5gm 199
10867 Albumin,Bovine,Protease-Free 10gm 198 50gm 100gm
10868 Albumin,Bovine,RIAGrade,pH7 10gm 198 50gm 100gm
10895 AlcoholDehydrogenase 75,000units 199
10906 Agar,Bacteriological,Ultrapure 1lb 190 5lb 25lb
10907 Agar,NobleAgar,Ultrapure 100gm 190 500gm 1kg
10925 ChickenIntestineAlkalinePhosphastase 1gm 206 5gm
11055 p-AminoBenzoicAcid(PABA) 200gm 199 500gm 1kg 5kg
11056 4-AminoAntipyrene 50gm 199 100gm
11079 L-a-Amino-N-ButyricAcid 1gm 199 5gm
11118 AEBSFHydrochloride 200mg 190 1gm
11251 AmmoniumAcetate,Ultrapure 500gm 199 1kg
11254 AmmoniumSulfate,Ultrapure 1kg 200 5kg
11259 Ampicillin,SodiumSalt,Ultrapure 5gm 200 25gm 100gm
11379 G-418Sulfate,Ultrapure 500mg 217 1gm 5gm
11388 Aprotinin,Ultrapure 10mg 200 25mg 100mg
11405 L-(+)-Arabinose 25gm 200 100gm 1kg
11406 L-Arabinose 25gm 200 100gm 1kg
11490 L-Arginine 100gm 200 500gm 1kg
11500 L-Arginine,Hydrochloride 100gm 201 500gm 1kg
11545 L-AscorbicAcid 1kg 201 5kg
11590 L-Asparagine,Anhydrous 100gm 201 500gm 1kg
Code Product Size Page Code Product Size Page
Product Code Index | Numerical
254 888-362-2447 | 216-765-5000 | usb.affymetrix.com
11595 L-Asparagine,Monohydrate 100gm 201 1kg 5kg
11620 L-AsparticAcid 100gm 201 1kg 5kg
11631 L-AsparticAcid,MagnesiumSalt,Dihydrate 100gm 201 1kg
11635 L-AsparticAcid,SodiumSalt,Monohydrate 1kg 201
11681 Streptavidin,Ultrapure 5mg 242 10mg
11687 Streptavidin-AlkalinePhosphataseConjugate 250µl 242 1,500µl
11805 Bacitracin 5gm 202 25gm
12091 Bicine 100gm 202 1kg
12110 Bilirubin 1gm 202 5gm
12112 Bis-Tris,Ultrapure 100gm 202 500gm 1kg
12355 5-Bromo-4-Chloro-3-IndolylBromocresolGreen, 5gm 203 SodiumSalt,Ultrapure 25gm
12370 BromophenolBlue,SodiumSalt,Ultrapure 5gm 204 10gm
12387 5-Bromo-4-Chloro-3-5-Bromo-4-Chloro-3-Indolyl 100mg 203 Phosphate,DisodiumSalt,Ultrapure 500mg 1gm
12488 CacodylicAcid,SodiumSaltTrihydrate 100gm 204 500gm
12520 Caffeine,Anhydrous,Ultrapure 100gm 204 1kg
12531 CalciumChloride,Dihydrate 500gm 204 2.5kg
12550 D-CalciumPantothenate 100gm 204 500gm 1kg
12678 Carbenicillin,DisodiumSalt 1gm 205 5gm
12758 CarboxypeptidaseY 1mg 205 10mg
12840 Casein,Hammarsten,Ultrapure 100gm 205 1lb 5lb 25lb
12845 Casein(HighNitrogen) 5lb 205 25lb 100lb
12855 CaseinHydrolysate,EnzymaticDigest,Ultrapure 1lb 205 5lb 25lb
12865 Casein,Sodium 1lb 205 5lb
12866 Casein-Vitafree,Ultrapure 5lb 205 25lb
12885 Catalase 500mg 205 1gm 5gm 10gm
13285 Cellulase 100gm 205
13353 CetylTrimethylAmmoniumBromide 100gm 206 500gm 1kg
13410 CholineChloride 200gm 207 1kg 5lb 25lb
13580 Cholesterol,ReagentGrade 5gm 206 100gm
13610 Cholesterol 100gm 206 1kg
13729 CitricAcid,Anhydrous 500gm 207 1kg
13735 CitricAcid,TrisodiumSalt,Dihydrate,Ultrapure 1kg 207 5kg
13765 Cocarboxylase 25gm 207 100gm
13785 CoenzymeA,TrilithiumSalt,Dihydrate 100mg 207 250mg 500mg 1gm
13787 CoenzymeA,FreeAcid,Trihydrate 50mg 207 100mg 250mg 500mg 1gm
13813 Collagen 10ml 207 25ml 100ml
13820 Collagenase,TypeI 100mg 207 1gm 5gm
13911 CRCLowerFilters,10µmporesize 1pk 208
13912 CRCLowerFilters,35µmporesize 1pk 208
13913 CRCLowerFilters,90µmporesize 1pk 208
13914 CRCUpperFilters,10µmporesize 1pk 208
13915 CreatineKinase 100mg 208 1gm
13918 CRCUpperFilters,35µmporesize 1pk 208
13919 CRCUpperFilters,90µmporesize 1pk 208
13920 CreatinePhosphate,DisodiumSalt,Tetrahydrate 5gm 208 25gm
13928 CompactReactionColumns 1pk 207
13929 CompactReactionColumns,withoutscrewonlids 1pk 208
13979 a-Cyclodextrin 25gm 208
Code Product Size Page Code Product Size Page
Product Code Index | Numerical
255For bulk or alternate pack sizes, email us at [email protected].
14035 L-Cysteine,Hydrochloride,Monohydrate 25gm 208 100gm 1kg 5kg
14050 L-Cystine 50gm 208 100gm 1kg 5kg
14060 Cytidine,FreeBase 25gm 208 100gm
14102 CytochromeC 100mg 209 500mg 1gm
14121 Cytidine-5’-Triphosphate,DisodiumSalt,Dihydrate 100mg 209 500mg 1gm
14235 2’-Deoxyadenosine 5gm 209 25gm
14284 2’-Deoxycytidine,Hydrochloride,Synthetic 1gm 210 5gm 10gm 100gm
14300 2’-Deoxyguanosine 5gm 210 25gm 100gm
14304 2’-Deoxyguanosine,MonohydrateSynthetic 1gm 210 5gm 10gm 100gm 1kg
14340 DNaseI 10mg 213 20mg
14365 DNaseI,Lyophilized 5mg 72
14367 DNaseI,Solution 100units 72 500units
14375 DNA,CalfThymus 1gm 128 5gm
14377 DNA,FishSperm,SodiumSalt 100gm 128 1kg
14380 DNA,E. coli 10mg 128
14394 DNA-Cellulose,Double-Stranded 5gm 124 10gm
14395 DNA-Cellulose,Single-Stranded 5gm 124 10gm
14405 DNA,FishSperm 100gm 128 1kg
14420 2-Deoxy-D-Ribose,Ultrapure 5gm 211 25gm 100gm
14489 DextranSulfate,SodiumSalt 10gm 211 50gm 100gm
14495 Dextran,Ultrapure 100gm 211 1kg
14535 D-(+)-Dextrose,Anhydrous 5lb 211 25lb 100lb
14536 D-(+)-Dextrose,Monohydrate 10kg 212
14564 DAPI 10mg 209 50mg
14615 Diaphorase 600units 212 1,800units
14710 DiethylPyrocarbonate,Ultrapure 25ml 212 100ml
14765 DihydrostreptomycinSulfate 100gm 213
14850 Dimedone 100gm 213 1kg
14868 p-DimethylaminoBenzaldehyde 100gm 213 500gm
14872 p-DimethylaminoCinnamaldehyde 25gm 213
15335 b-NicotinamideAdenineDinucleotide 1gm 229 5gm 10gm 25gm 100gm
15345 b-NicotinamideAdenineDinucleotide-Reduced, 1gm 230 Disodium,Trihydrate 5gm 10gm 25gm
15385 Dithioerythritol,Ultrapure 5gm 213 25gm 100gm
15395 Dithiothreitol 5gm 213 25gm 100gm
15397 Dithiothreitol,Ultrapure 1gm 213 5gm 25gm 100gm
15460 Albumin,Egg 1lb 199 5lb 25lb
15475 Elastase 5mg 214 10mg 20mg 100mg
15515 Enolase 10,000units 214
15562 Ergocalciferol 5gm 214 25gm 100gm
15590 Esculin 100gm 214
15694 EthylenediaminetetraaceticAcid,(EDTA),0.5M 100ml 215 Solution,Ultrapure 500ml
15697 EthylenediaminetetraaceticAcid,(EDTA)Disodium 1kg 215 Salt,Dihydrate
15699 EthylenediaminetetraaceticAcid,(EDTA)Disodium 500gm 215 Salt,Dihydrate 5kg
15700 EthyleneDiamineTetraaceticAcid,(EDTA)Tetrasodium 1kg 215 Salt,Dihydrate,Ultrapure
Code Product Size Page Code Product Size Page
Product Code Index | Numerical
256 888-362-2447 | 216-765-5000 | usb.affymetrix.com
15701 EthylenediaminetetraaceticAcid,(EDTA)Disodium 100gm 215 Salt,Dihydrate,Ultrapure 500gm 1kg
15703 EthyleneGlycol-O,O’-Bis(2Amino-ethyl)-N,N,N’,N’,- 10gm 215 TetraaceticAcid(EGTA),Ultrapure 50gm 100gm
15762 FetalBovineSerum(FBS) 500mI 215
15785 FlavinAdenineDinucleotide,DisodiumSalt,Hydrate 250mg 216 500mg 1gm
15880 FolicAcid 25gm 216 100gm 1kg
15929 D-Fructose-6-Phosphate,DisodiumSalt 1gm 216 5gm
15995 D-(+)-Galactose 500gm 217 1kg 5kg
16021 g-Globulins,FractionII 10gm 217 25gm
16051 GentamicinSulfate 1gm 217 5gm 25gm
16146 GlucoseOxidase 50,000units 217 250,000units 1mu
16171 Glucose-6-PhosphateDehydrogenase 1,000units 217 Source:Leuconostoc mesenteroides 5,000units 10,000units
16176 Glucose-6-PhosphateDehydrogenase 1,000units 218 Source:Yeast 5,000units 10,000units
16185 D-Glucose-6-Phosphate,DisodiumSalt,Dihydrate 5gm 217 10gm
16215 L-GlutamicAcid 1kg 218 5kg 25kg
16245 L-GlutamicAcid,MonosodiumSalt,Monohydrate 5kg 218
16250 Glucose-6-PhosphateGlutamateDehydrogenase 100mg 218 500mg 1gm
16285 L-Glutamine 100gm 218 250gm 500gm 1kg
16315 Glutathione-Reduced 25gm 218 100gm
16320 Glutathione-Oxidized 250mg 218 1gm
16372 Glycine 2.5kg 220
16374 Glycerol,Ultrapure 500ml 219 1L
16405 Glycine 1kg 220 5kg 25kg
16407 Glycine,Ultrapure 500gm 219 1kg 5kg
16445 Glycogen,Ultrapure 5gm 220 10gm 25gm
16505 Glycyl-Glycine 100gm 220 1kg
16785 Guanosine-5’-Diphosphate,DisodiumSalt 100mg 221 500mg 1gm
16800 Guanosine-5’-Triphosphate,DisodiumSalt 25mg 221 100mg 500mg 1gm
16880 Hemin 10gm 221 50gm
16920 Heparin,SodiumSalt 50,000units 221 100,000units 500,000units 1mu
16924 HEPES,1MSolution,pH7.3,Ultrapure 100ml 222 500ml 1L
16926 HEPES,Ultrapure 100gm 222 250gm 1kg 5kg
16928 HEPES,SodiumSalt 100gm 222 1kg
16965 Hexokinase 1,000units 222 10,000units 25,000units 50,000units
17070 L-Histidine,FreeBase 100gm 222 1kg
17080 L-Histidine,Monohydrochloride,Monohydrate 100gm 222 1kg 5kg
17525 Imidazole,Ultrapure 100gm 223 500gm 1kg 5kg
17798 IsocitrateDehydrogenase,(NADP+Specific), 150units 223 Recombinant 600units
17820 DL-Isoleucine 100gm 223 500gm 1kg
17825 L-Isoleucine 10gm 223 100gm 500gm 1kg
17886 Isopropyl-b-D-Thiogalactopyranoside(IPTG),Dioxane 1gm 223 Free,Ultrapure 5gm 25gm 100gm
17924 KanamycinSulfate 5gm 223 25gm
Code Product Size Page Code Product Size Page
Product Code Index | Numerical
257For bulk or alternate pack sizes, email us at [email protected].
17950 a-KetoglutaricAcid,FreeAcid 100gm 224 500gm 1kg
17955 a-KetoglutaricAcid,DisodiumSalt,Dihydrate 5gm 224 100gm 1kg
18160 L-(+)-LacticAcid,LithiumSalt 100gm 224
18177 Lactoferrin 100mg 224 500mg 1gm
18185 D-(+)-Lactose,Monohydrate 5kg 224
18218 N-DodecylTrimethylammoniumBromide 10gm 214 50gm 250gm
18220 SodiumDodecylSulfate(SDS) 500gm 240 1kg 5kg
18245 Lecithin 500gm 224 1kg
18270 DL-Leucine 100gm 225 1kg
18280 L-Leucine 100gm 225 1kg 5kg
18413 Leupeptin,Ultrapure 5mg 225 25mg 100mg
18425 b-D-Fructose 1kg 216
18505 DL-a-LipoicAcid 25gm 225 50gm 100gm
18522 DL-a-LipoicAcid 50gm 225 100gm 1kg
18585 L-Lysine,Hydrochloride 1kg 226 5kg 10kg
18641 MagnesiumChloride,Hexahydrate,Ultrapure 500gm 226 1kg
18645 Lysozyme,Ultrapure 5gm 226 10gm 25gm 100gm
18670 MalateDehydrogenase,Lyophilized 25,000units 226 100,000units
18685 MaleicAcid 1kg 226
18695 DL-MalicAcid 1kg 226
18700 L-MalicAcid 500gm 227 1kg
18724 Maltose,Monohydrate 500gm 227 1kg
18725 D-(+)-Maltose,Monohydrate 1kg 227 5kg
18755 Mannitol 500gm 227 1kg 5kg
18775 D-(+)-Mannose 100gm 227 500gm
18886 MES,Monohydrate,Ultrapure 100gm 227 1kg
18888 MES,Anhydrous 100gm 227 250gm 1kg
18893 MES,SodiumSalt 100gm 227 250gm 1kg
19184 5-Methyl-2’-Deoxycytidine-5’-Monophosphate, 100mg 228 DisodiumSalt
19220 MethyleneBlue,Trihydrate 100gm 228 500gm
19256 MOPS,Ultrapure 100gm 228 1kg
19265 MTT,Ultrapure 500mg 228 1gm 5gm 10gm
19391 a-NaphthylPhosphate,MonosodiumSalt 25gm 229
19430 Neocuproine,Hydrochloride 10gm 229 25gm 100gm 1kg
19435 NeomycinSulfate,Ultrapure 10gm 229 25gm
19438 b-NicotinamideAdenineDinucleotide,LithiumSalt, 1gm 229 Dihydrate 5gm
19439 b-NicotinamideAdenineDinucleotide,Monosodium 5gm 230 Salt
19465 NeutralRed 25gm 229 100gm 1kg
19480 Niacinamide 1kg 229 10kg
19535 p-NitroBlueTetrazoliumChloride,Ultrapure 250mg 230 1gm 5gm 10gm
19594 4-Nitrophenyl-b-D-Glucopyranoside 1gm 231 5gm
19629 SSC,20XSolution,Ultrapure 500ml 241 1L 5L
19817 Oligop(dT)12-18SodiumSalt 5units129,232 25units 100units
19880 Pancreatin1X 200gm 232
19943 ParaformaldehydeSolution,4%inPBS 1L 232
19957 PolyethyleneGlycol400 500ml 235 1L
Code Product Size Page Code Product Size Page
Product Code Index | Numerical
258 888-362-2447 | 216-765-5000 | usb.affymetrix.com
19959 PolyethyleneGlycol8000,Flakes,Ultrapure 500gm 235 1kg 5kg
19966 PolyethyleneGlycol8000,Powder 500gm 235 1kg 5kg
19972 PolyethyleneGlycol6000,Flakes 1kg 235 5kg
19985 PenicillinG,Potassium 1kg 232
20010 Pepsin 1gm 232 5gm 10gm 25gm
20015 Pepsin1:10,000 100gm 233 500gm 1kg
20037 PepstatinA,Ultrapure 5mg 233 25mg
20048 BacteriologicalPeptone,Ultrapure 1lb 202 5lb 25lb
20075 Phenolphthalein 500gm 234 1kg
20085 PhenolRed,SodiumSalt 10gm 234 25gm 50gm
20100 DL-Phenylalanine 100gm 234 1kg
20105 L-Phenylalanine 100gm 234 250gm 1kg 5kg
20203 PhenylmethylSulfonylFluoride,Ultrapure 5gm 234 25gm 100gm
20227 PotassiumPhosphate,Monobasic,Anhydrous, 500gm 236 Ultrapure 1kg
20228 PotassiumPhosphate,Monobasic 1kg 236 10kg
20229 SodiumPhosphate,Monobasic 1kg 240 5kg
20232 SodiumPhosphate,Dibasic,Heptahydrate,Ultrapure 500gm 240 1kg
20233 SodiumPhosphate,Monobasic,Monohydrate, 500gm 241 Ultrapure 1kg
20240Y PhosphodiesteraseI 100units 81
20274 PotassiumPhosphate,Dibasic,Ultrapure 500gm 236 1kg 2.5kg
20276 PotassiumPhosphate,Dibasic 1kg 236 10kg
20426 PIPES 100gm 208 1kg
20428 PIPES,DisodiumSalt 10gm 234 100gm 1kg
20450 PolyanetholesulfonicAcid,SodiumSalt 5gm 234 100gm
20539 Poly(dI-dC)•Poly(dI-dC)SodiumSalt 5units 235 25units 100units
20551 PolymixinBSulfate 10mu 235 100mu
20598 PotassiumChloride,Ultrapure 500gm 236 1kg 5kg
20611 PVP-K-30 1kg 237
20795 ProtamineSulfate 25gm 237 100gm
20995 PyruvicAcid,SodiumSalt 10gm 237 25gm 100gm 500gm
21026 DeoxycholicAcid,SodiumSalt 25gm 210 100gm
21060 D-(+)-Raffinose,Pentahydrate 100gm 237 500gm 1kg
21145 Riboflavin 25gm 238 100gm 500gm 1kg
21185 RNA,FreeAcid 100gm 238 1kg
21190 RNA,SodiumSalt 25gm 239 100gm 1kg
21195 RNaseA,Lyophilized 100mg 239 1gm 5gm
21220 D-(–)-Ribose 10gm 238 100gm
21245 T4RNALigase 600units 71 2,500units
21246 Rifampin 1gm 238 5gm
2141Y KlenowDNAPolymeraseI 300units 872141Z 1,500units
21486 DL-Serine(Non-Animal) 100gm 239 1kg
21610 SodiumAzide,Ultrapure 100gm 239 500gm
21616 SodiumCarbonate,Anhydrous 500gm 239 2.5kg
21618 SodiumChloride,Ultrapure 500gm 239 1kg 5kg 10kg
Code Product Size Page Code Product Size Page
Product Code Index | Numerical
259For bulk or alternate pack sizes, email us at [email protected].
21653 N-LauroylSarcosine,SodiumSalt,Ultrapure 50gm 224 100gm
21655 b-Glycerophosphate,SodiumSalt,Pentahydrate 100gm 219 500gm 1kg
21660 SodiumPhosphate,DibasicAnhydrous 500gm 240 2.5kg
21665 SodiumSelenite 100gm 241 500gm
21695 Starch-Soluble,Ultrapure 1kg 242
21710 D-Sorbitol,Ultrapure 500gm 241 1kg 5kg
21755 Spermine,Tetrahydrochloride 5gm 241
21760 Spermidine,Trihydrochloride,Ultrapure 1gm 241 5gm
21865 StreptomycinSulfate,Ultrapure 10gm 242 100gm 1kg
21900 SuccinicAcid,FreeAcid 500gm 242 1kg 5kg
21910 SuccinicAcid,DisodiumSalt,Hexahydrate 500gm 242
21930 Sucrose,Crystalline 1kg 243 5lb 100lb
21931 Sucrose 500gm 243 2.5kg
21938 Sucrose,Ultrapure 1kg 243 5kg
21985 SulfanilicAcid 500gm 243
22086 TES,Ultrapure 100gm 245 500gm 1kg
22105 Tetracycline,Hydrochloride,Ultrapure 25gm 245 100gm 1kg
22124 TetramethylammoniumChloride,Ultrapure 1kg 245
22126 3,3’,5,5’-Tetramethylbenzidine 1gm 245 5gm
22170 Thiamine,Hydrochloride 100gm 245 500gm 1kg
22215 Thimerosal 25gm 246 100gm
22290 L-Threonine 100gm 246 1kg
22292 ThrombinHuman 1,000units 246
22293 Thrombin 25,000units 246 100,000units
22515 a,a-Trehalose,Dihydrate 100gm 246
22561 Tricine,Ultrapure 100gm 246 500gm 1kg
22565 Triethanolamine,Hydrochloride 250gm 246 1kg
22631 2,3,5-TriphenylTetrazoliumChloride 10gm 246 25gm 50gm 100gm 500gm 1kg
22637 Tris/HCl,1MSolution,pH7.0,Ultrapure 100ml 248 500ml 1L
22638 Tris/HCl,1MSolution,pH8.0,Ultrapure 100ml 248 500ml 1L
22639 Tris/HCl,1MSolution,pH7.5,Ultrapure 100ml 248 500ml 1L
22650 b-NicotinamideAdenineDinucleotidePhosphate- 25mg 230 Reduced,Tetrasodium,Tetrahydrate 100mg 250mg 500mg 1gm
22655 b-NicotinamideAdenineDinucleotidePhosphate, 100mg 230 Monosodium,Tetrahydrate 250mg 500mg 1gm 5gm
22660 2,4,6-Tripyridyl-s-Triazine(TPTZ) 5gm 247 25gm 100gm
22674 Tris,HighPurityGrade 500gm 247 1kg 5kg
22675 Tris,ReagentGrade 500gm 247 1kg
22676 Tris,Hydrochloride,Ultrapure 500gm 247 1kg 5kg
22710 Trypsin1:250 100gm 248 1kg
22720 Trypsin,Ultrapure 1gm 248 5gm 10gm
22725 Trypsin(TPCKTreated) 100mg 248 250mg
22765 L-Tryptophan 100gm 248 1kg
22885 Tyrosinase 100,000units 248 500,000units
22910 L-Tyrosine 100gm 248 1kg 5kg
23020 Uracil 100gm 249 1kg
Code Product Size Page Code Product Size Page
Product Code Index | Numerical
260 888-362-2447 | 216-765-5000 | usb.affymetrix.com
23036 Urea 1kg 249 5kg
23075 UricAcid 100gm 249 500gm 1kg
23130 Uridine 1gm 249 10gm 100gm
23160 Uridine-5’-Triphosphate,TrisodiumSalt 100mg 249 1gm
23261 Vancomycin,Hydrochloride 100mg 250 250mg
23360 VitaminB12 1gm 250
23445 VitaminK1 250mg 250 1gm 5gm 10gm
23457 Wright’sStain 5gm 250 10gm 25gm 100gm
23465 XanthineMonosodiumSalt,Monohydrate 5gm 250 10gm 25gm 100gm
23513 XyleneCyanolFF,Ultrapure 25gm 251
23520 D-(+)-Xylose 500gm 251 1kg
23540 Yeast,Bakers 1lb 251 5lb 25lb
23545 YeastExtractPowder,LowDust 1kg 251 5kg
23546 Yeast,Brewers 5lb 251 25lb 100lb
23547 YeastExtractPowder,Ultrapure 1kg 251
23550 YeastHydrolysate,Enzymatic,Ultrapure 5lb 251 25lb 50lb 100lb
23551 Yeast-Torula 5lb 251 25lb 100lb
23660 Chloramphenicol,Ultrapure 25gm 206 100gm 1kg
25110 LeptomycinB 1µg 224
26004 Thioredoxin,Human,Recombinant 100µg 246
26200 5-Bromo-4-Chloro-3-Indolyl-a-D-Galactopyranoside, 100mg 203 Ultrapure 500mg
27-0323-01 RNaseI‘A’,Powder 100mg 79,163
27-0330-02 RNaseI 1gm 79,163
32800 Acrylamide 25gm 188 100gm 500gm 1kg
32802 Agarose-LE,Ultrapure 25gm 194 100gm 250gm 500gm 1kg
32803 Agarose-ME,Ultrapure 25gm 192 100gm 250gm 500gm
32804 Agarose-HE,Ultrapure 100gm 192 250gm 500gm
32812 BrilliantBlueG,Ultrapure 25gm 203 100gm
32813 EthidiumBromide,Ultrapure 1gm 214 5gm 25gm
32816 LithiumDodecylSulfate,Ultrapure 25gm 225 100gm
32819 PonceauS,SodiumSalt,Ultrapure 50gm 236
32821 Agarose-UltraLowMelt,Ultrapure 10gm 197 25gm
32823 Starch,Hydrolyzed,Ultrapure 100gm 242 1kg
32826 BrilliantBlueR,Ultrapure 25gm 203 100gm
32829 Agarose-LowMeltSeparation≤1000bp,Genetic 25gm 197 PerformanceCertified,Ultrapure 100gm
32830 Agarose-LowMeltSeparation≥1000bp,Genetic 25gm 196 PerformanceCertified,Ultrapure 100gm
33230 4-Methylumbelliferyl-a-L-Iduronide,SodiumSalt 5mg 228
34007 GuanidineHydrochloride 1kg 221 5kg
34128 4-Nitrophenyl-N-Acetyl-b-D-Galactosaminide 100mg 230 1gm
34131 4-Nitrophenyl-a-L-Fucopyranoside 50mg 231 250mg 1gm
34133 4-Nitrophenyl-a-D-Galactopyranoside 500mg 231 1gm 5gm
34136 4-Nitrophenyl-b-D-Glucuronide 100mg 231 250mg 1gm
34138 4-Nitrophenyl-a-D-Maltoside 50mg 231 100mg
34140 4-Nitrophenyl-b-D-Mannopyranoside 100mg 231 500mg
34142 4-Nitrophenyl-b-D-Xylopyranoside 500mg 231 1gm
Code Product Size Page Code Product Size Page
Product Code Index | Numerical
261For bulk or alternate pack sizes, email us at [email protected].
70001Z T7RNAPolymerase,Highconcentration, 30,000units 92,161 >80units/µl
70005Y T4DNALigase,Standardconcentration,1unit/µl 100units 7070005X 500units70005 2,000units
70010Y DNAPolymeraseI 500units 87
70017Z T7DNAPolymerase 250units 8870017 1,000units
70020Y E. coliDNALigase 200units 7070020Z 1,000units
70023Y ExonucleaseIII 5,000units 7770023Z 25,000units
70025Y T7Gene6Exonuclease 2,000units 7870025Z 10,000units
70028Y RecAProtein 200µg 6570028Z 1,000µg
70029Y T4Gene32Protein,Standardconcentration,5µg/µl 100µg 6670029Z 500µg
70031Y T4PolynucleotideKinase(PNK) 500units 6970031Z 1,000units70031X 2,500units
70032Y Single-StrandedDNABindingProtein(SSB) 100µg 6570032Z 500µg
70034Y CalfIntestinalAlkalinePhosphatase(CIAP) 1,500units 82
70041Y AMVReverseTranscriptase 200units 90,16270041Z 1,000units
70042X T4DNALigase,Highconcentration,10units/µl 500units 7070042 2,000units
70047Y T7RNAPolymerase,Standardconcentration, 6,000units 92,161 20units/µl
70051Y T3RNAPolymerase,Standardconcentration, 1,000units 92,16070051Z 20units/µl 5,000units
70052 TthDNAPolymerase 200units 89 1,000units
70054Y RNaseH 250units 80,16370054Z 1,000units
70057Y Exonuclease-FreeKlenow 125units 8670057Z 750units70057 2,500units
70065 7-deaza-2’-Deoxyguanosine-5’-Triphosphate, 2µmol 60,210 LithiumSalt,10mMSolution
70073Z ExonucleaseI,Standardconcentration,10units/µl 2,500units 7670073X 5,000units
70076 DNA,FishSperm,Activated 500µg 128 2mg
70082Y ExonucleaseVII 200units 7770082Z 1,000units
70140 SequenaseQuickDenaturePlasmidDNA 100reactions 177 SequencingKit
70170 SequenasePCRProductSequencingKit 100reactions 176
70175 SequenaseVersion2.0DNAPolymerasewith 325units 85 Pyrophosphatase
70194Y RNaseA,MolecularBiologyGrade,Solution 1mg 80,16370194Z 5mg
70196Y MicrococcalNuclease 15,000units 81
70204 T3RNAPolymerase,Highconcentration, 1,000units 92,160 200units/µl 5,000units
70440 T4RNALigase2 200units 71 1,000units
70454 TBEBuffer,10XReady-MixedPowder,Ultrapure 6x200ml 243
70468 Denhardt’sSolution,50X,Ultrapure 50ml 209
70704 M13mp18,Single-Stranded,0.2µg/ul 50µl 125 50µg
70724 StopSolution,Ultrapure 1,200µl 242
70726 Dithiothreitol(DTT),0.1MSolution,Ultrapure 150µl 213 1ml 5ml
70770 SequenaseVersion2.0DNASequencingKit 100reactions 175
70775Y SequenaseVersion2.0DNAPolymerase 200units 88,17470775Z 1,000units
70783 Water,RNase-Free,DEPCTreated,Ultrapure 25ml 250 10x1ml 100ml 500ml 1L
70796 DextranSulfate,SodiumSalt,Ultrapure 10gm 211 50gm
70995 PCRProductPre-SequencingKit 100reactions 5270996 500reactions70997 2,000reactions
71155 HotStart-ITFideliTaqDNAPolymerase 50units 17,25 250units 1,000units 5,000units
71156 HotStart-ITFideliTaqPCRMasterMix(2X) 25reactions 18 100reactions 500reactions
71160 TaqDNAPolymerase 50units 11 250units 1,000units 5,000units
71161 TaqPCRKit 100reactions(100µl/rctn) 12
71162 TaqPCRMasterMix(2X) 100reactions 12
71170 VeriQuestTaqDNAPolymerase 50units 26 250units 1,000units 5,000units
71180 FideliTaqDNAPolymerase 50units 15 250units 1,000units 5x250units 5,000units
71182 FideliTaqPCRMasterMix(2X) 100reactions 16
Code Product Size Page Code Product Size Page
Product Code Index | Numerical
262 888-362-2447 | 216-765-5000 | usb.affymetrix.com
71185 FideliTaqRT-PCRMasterMix(2X) 100reactions 46
71190 ROXPassiveReferenceDye 50units 13
71191 RubyTaqPCRMasterMix(2X) 100reactions 14 500reactions
71194 HotStart-ITBindingProtein 400μg 21
71195 HotStart-ITTaqDNAPolymerase 50units 19 250units 1,000units 5x250units 5,000units
71196 HotStart-ITTaqPCRMasterMix(2X) 25reactions 20 100reactions 500reactions
71199 MagniTaqMultiplexPCRMasterMix(2X) 25reactions 22 100reactions
71200 T3PromoterPrimer 250pmol 178
71547 Oligop(dT)Cellulose,Ultrapure 1gm 125
71570 RNaseInhibitor(HumanPlacenta) 5,000units 94,164
71571 RNaseInhibitor(Recombinant) 5,000units 94,164
71600 MineralOil,Ultrapure 10ml 228 25ml 1L
71706 M13mp18DNA,Single-Stranded,1.0µg/µl 50µg 125 200µg
71786 Water,Nuclease-Free,Ultrapure 10x1ml 250 100ml 500ml 1L 5L
71960 Uracil-DNAGlycosylase,E. coli 400units 67 2,000units
72020 Exonuclease-FreeKlenow,TrisBuffer 750units 86 2,500units
72033 TerminalDeoxynucleotidylTransferase, 500units 93 Recombinant(rTdT) 2,500units
72073 ExonucleaseI,Highconcentration,20units/µl 5,000units 76
74015 TAEBuffer,50XSolution,Ultrapure 100ml 243
74029Y T4Gene32Protein,Highconcentration,≥10µg/µl 300µg 6674029Z 1,000µg
74225Y Poly(A)Polymerase,Yeast 10,000units 91,15974225Z 25,000units
75592 SuperSAPShrimpAlkalinePhosphatase 50reactions 84
75600 VeriQuestSYBRGreenqPCRMaster 40reactions(1ml) 27 Mix(2X) 200reactions(5ml) 400reactions(2x5ml) 1,000reactions(5x5ml) 2,000reactions(10x5ml)
75650 VeriQuestProbeqPCRMasterMix(2X) 40reactions(1ml) 33 200reactions(5ml) 400reactions(2x5ml) 1,000reactions(5x5ml) 2,000reactions(10x5ml)
75660 VeriQuestProbeqPCRMasterMix, 40reactions(1ml) 35 NoReferenceDye(2X) 200reactions(5ml) 400reactions(2x5ml) 1,000reactions(5x5ml) 2,000reactions(10x5ml)
75665 VeriQuestSYBRGreenqPCRMaster 40reactions(1ml) 29 MixwithFluorescein(2X) 200reactions(5ml) 400reactions(2x5ml) 1,000reactions(5x5ml) 2,000reactions(10x5ml)
75675 VeriQuestFastSYBRGreenqPCR 100reactions(1ml) 31 MasterMixwithFluorescein(2X) 500reactions(5ml) 1,000reactions(2x5ml) 2,500reactions(5x5ml) 5,000reactions(10x5ml)
75680 VeriQuestFastProbeqPCRMasterMix(2X) 100reactions(1ml) 36 500reactions(5ml) 1,000reactions(2x5ml) 2,500reactions(5x5ml) 5,000reactions(10x5ml)
75685 VeriQuestFastProbeqPCRMaster 100reactions(1ml) 37 Mix,NoReferenceDye(2X) 500reactions(5ml) 1,000reactions(2x5ml) 2,500reactions(5x5ml) 5,000reactions(10x5ml)
75690 VeriQuestFastSYBRGreenqPCR 100reactions(1ml) 30 MasterMix(2X) 500reactions(5ml) 1,000reactions(2x5ml) 2,500reactions(5x5ml) 5,000reactions(10x5ml)
75700 VeriQuestProbeOne-StepqRT-PCR 40reactions(1ml) 42 MasterMix(2X) 200reactions(5ml)
75705 VeriQuestSYBRGreenOne-StepqRT-PCR 40reactions(1ml) 40 MasterMix(2X) 200reactions(5ml)
75710 VeriQuestProbeOne-StepqRT-PCR 40reactions(1ml) 43 MasterMix,NoReferenceDye(2X) 200reactions(5ml)
75715 VeriQuestSYBRGreenOne-StepqRT-PCR 40reactions(1ml) 41 MasterMixwithFluorescein(2X) 200reactions(5ml)
75760 HotStart-ITSYBRGreenqPCRMasterMix 100reactions 32 withUDG(2X) 500reactions
75762 HotStart-ITSYBRGreenqPCRMasterMix(2X) 100reactions 32 500reactions
75764 HotStart-ITProbeqPCRMasterMixwith 100reactions 38 UDG(2X) 500reactions
75766 HotStart-ITProbeqPCRMasterMix(2X) 100reactions 38 500reactions
75767 FluoresceinPassiveReferenceDye 500μl 53
75768 RT-PCRMasterMix(2X) 500μl 53
75770 HotStart-ITSYBRGreenOne-StepqRT-PCRMaster 100reactions 44 MixKit 500reactions
75772 HotStart-ITProbeOne-StepqRT-PCRMaster 100reactions 44 MixKit 500reactions
75780 First-StrandcDNASynthesisKitfor 50reactions(20µl) 39 Real-TimePCR
75790 TEBuffer,1XSolution,pH7.0 100ml 244 500ml
Code Product Size Page Code Product Size Page
Product Code Index | Numerical
263For bulk or alternate pack sizes, email us at [email protected].
75791 TEBuffer,1XSolution,pH7.0,LowEDTA 100ml 244 500ml
75792 TEBuffer,1XSolution,pH8.0 100ml 244 500ml
75793 TEBuffer,1XSolution,pH8.0,LowEDTA 100ml 244 500ml
75816 EthidiumBromideDrops,Ultrapure 5ml 214
75817 Agarose-Separation≥500bp,GeneticPerformance 25gm 193 Certified,Ultrapure 100gm 250gm 500gm
75818 GuanidineThiocyanate,Ultrapure 100gm 221 250gm 1kg
75819 SodiumDodecylSulfate(SDS),Ultrapure 100gm 240 1kg
75820 Acrylamide,Ultrapure 100gm 188 500gm 1kg
75821 N,N’-Methylene-bis-Acrylamide,Ultrapure 25gm 228 50gm 100gm 1kg
75822 CesiumChloride,Ultrapure 5gm 206 25gm 100gm 500gm 1kg
75823 GuanidineHydrochloride,Ultrapure 100gm 220 500gm
75824 Taurine,Ultrapure 1kg 243
75825 Tris,Ultrapure 500gm 247 1kg 5kg 10kg
75826 Urea,Ultrapure 1kg 249 5kg
75827 GlycerolTolerantGel(GTG)Buffer,20XSolution, 1L 219 Ultrapure
75828 Formamide,Ultrapure 500gm 216 1kg
75829 Phenol,pH8.0,Equilibrated,Ultrapure 100ml 233 400ml
75830 Phenol,Ultrapure 500gm 233
75831 Phenol:Chloroform:IsoamylAlcohol(25:24:1), 100ml 233 Ultrapure 400ml
75832 SodiumDodecylSulfate(SDS),20%Solution,Ultrapure 100ml 240 500ml
75834 TEBuffer,50XSolution,Ultrapure 100ml 244 500ml
75848 RapidGel40%LiquidAcrylamideStockSolution, 500ml 237 Ultrapure
75851 LBAgar,Ready-MadePowder 250gm 224 1kg
75852 LBBroth,Ready-MadePowder 250gm 224 1kg
75853 LuriaAgar,Ready-MadePowder 250gm 225 1kg
75854 LuriaBroth,Ready-MadePowder 250gm 225 1kg
75855 NZCYMBroth,Ready-MadePowder 1kg 232
75856 TerrificBroth,Ready-MadePowder 250gm 245 1kg
75861 RapidGel-XL6%LiquidAcrylamideDNASequencing 500ml 238 Gel,TBEFormulation,Ultrapure
75864 2XYTBroth 1kg 251
75888 SodiumChloride(NaCl),5MSolution,Ultrapure 100ml 239 500ml 1L
75889 PBS,10XSolution,pH7.4,Ultrapure 100ml 232 1L 5L
75890 SSPE,20XSolution,Ultrapure 1L 241 5L
75891 TBEBuffer,5XSolution,Ultrapure 1L 243 5L
75892 TBS,20XSolution,pH7.4,Ultrapure 100ml 244 1L 5L
75893 TEBuffer,1XSolution,Ultrapure 10x1ml 244 100ml 500ml
75894 Tris-Glycine(TG)Buffer,10XSolution,Ultrapure 1L 247 5L
75895 Tris-Glycine-SDS(TGS)Buffer,10XSolution,Ultrapure 1L 247 5L
75896 PotassiumChloride(KCl),2MSolution,Ultrapure 100ml 236
75897 SodiumAcetate(NaOAc),3MSolution,pH5.5, 100ml 239 Ultrapure 500ml
75901 AmmoniumAcetate(NH4OAc),5MSolution,Ultrapure 100ml 199 500ml
75902 LithiumChloride(LiCl),7.5MSolution,Ultrapure 100ml 225
75904 TAEBuffer,10XSolution,Ultrapure 1L 243 5L
76225 ProteinaseKSolution 100mg 137
76230Y ProteinaseK 100mg137,23776230Z 500mg76230 1gm(2x500mg)
76320 N,N,N’,N’-Tetramethylethylenediamine,Ultrapure 100ml 245 100gm 500gm
76322 AmmoniumPersulfate(APS),Ultrapure 10gm 200 100gm 1kg
76324 BoricAcid,Ultrapure 500gm 202 1kg
Code Product Size Page Code Product Size Page
Product Code Index | Numerical
264 888-362-2447 | 216-765-5000 | usb.affymetrix.com
76410 LowMolecularWeightMarker,10-100nt 100µl132,165
76455 Poly(A)Tail-LengthAssayKit 1kit 158
76632 pUC19DNA,0.2µg/µl 10µg 129 50µg 200µg
76634 pUC19DNA,1.0µg/µl 50µg 129 200µg
76710 PCRMarkers,50-2000bp 250µl 130
76712 DNALadder,100bp 500µl 130
76714 DNALadder,1kbPlus 500µl 131
76715 DNALoadingBuffer(OXG),6X 1ml 130 5ml
76720 DNALoadingBuffer(BXF),6X 1ml 131
76740 ProteinMarkers,10-225kDa 500µl 136
77100 2’-Deoxynucleoside-5’-Triphosphates,SodiumSalt, 1pk 59,211 100mMSolutions,SetsofFour,Ultrapure 4x25µmol(250µl)eachdNTPperpack
77102 2’-Deoxyadenosine-5’-Triphosphate,Sodium 25µmol(250µl) 59,210 Salt,100mMSolution,Ultrapure 100µmol(1ml) 500µmol(5ml)
77104 2’-Deoxycytidine-5’-Triphosphate,SodiumSalt, 25µmol(250µl) 59,210 100mMSolution,Ultrapure 100µmol(1ml) 500µmol(5ml)
77106 2’-Deoxyguanosine-5’-Triphosphate,Sodium 25µmol(250µl) 59,210 Salt,100mMSolution,Ultrapure 100µmol(1ml) 500µmol(5ml)
77108 2’-Deoxythymidine-5’-Triphosphate,Sodium 25µmol(250µl) 59,211 Salt,100mMSolution,Ultrapure 100µmol(1ml) 500µmol(5ml)
77110 2’,3’-Dideoxyadenosine-5’-Triphosphate, 0.5µmol(50µl) 60,212 Ultrapure,10mMaqueoussolution,pH7.5 1µmol(100µl)
77112 2’,3’-Dideoxycytidine-5’-Triphosphate, 0.5µmol(50µl) 60,212 Ultrapure,10mMaqueoussolution,pH7.5 1µmol(100µl)
77116 2’,3’-Dideoxythymidine-5’-Triphosphate, 0.5µmol(50µl) 60,212 Ultrapure,10mMaqueoussolution,pH7.5 1µmol(100µl)
77119 PCRNucleotideMix,25mMSolution,Ultrapure 500µl 60,232
77125 2’,3’-Dideoxynucleoside-5’-Triphosphates,10mM 1pk 60,212 Solutions,SetsofFour,Ultrapure,4x1.0µmol(100µl) eachddNTPperpack
77126 2’,3’-Dideoxynucleoside-5’-Triphosphates,10mM 1pk 60,212 Solutions,SetsofFour,Ultrapure,4x0.5µmol(50µl) eachddNTPperpack
77128 2’-Deoxynucleoside-5’-Triphosphates,SodiumSalt, 1pk 59,211 100mMSolutions,SetsofFour,Ultrapure 4x500µmol(5ml)eachdNTPperpack
77206 2’-Deoxyuridine-5’-Triphosphate,SodiumSalt, 25µmol(250µl) 59,211 100mMSolution,Ultrapure 100µmol(1ml) 500µmol(5ml)
77212 PCRNucleotideMix,10mMSolution,Ultrapure 500µl 60,232 2x500µl
77241 Adenosine-5’-Triphosphate,100mMSolution, 25µmol(250µl) 61,189 Ultrapure
77243 Guanosine-5’-Triphosphate,SodiumSalt, 25µmol(250µl) 61,221 100mMSolution,Ultrapure
77245 Nucleoside-5’-Triphosphates(ATP,CTP,GTP,UTP), 1pk 61,231 SetofFour,100mMSolutions,Ultrapure 4x25µmol(250µl)eachNTPperpack
77328 2’-Deoxynucleoside-5’-Triphosphates,SodiumSalt, 1pk 59,211 100mMSolutions,SetsofFour,Ultrapure 4x100µmol(1ml)eachdNTPperpack
77330 PCRNucleotideMixwithdUTP,10mMSolution, 500µl 60,232 Ultrapure
77335 2’,3’-Dideoxynucleoside-5’-Triphosphates,100mM 1pk 60,212 Solutions,SetofFour,Ultrapure,4x4µmol(40µl) eachddNTPperpack
77405 Oligo(dT)12-18Primer,MW≅4500 100µl(2.5nmol)129,231
77500 TBSwithTween(TBST),20XSolution,pH7.4, 1L 244 Ultrapure 5L
77504 SodiumDodecylSulfate(SDS),10%Solution, 100ml 240 Ultrapure 500ml 1L
77507 Betaine,5MSolution,Ultrapure 1.5ml 202 5x1.5ml 10ml
77523 RapidRunAgaroseBuffer,20XSolution,Ultrapure 1L 238 5L
77524 RapidRunLoadingDye,Ultrapure 1ml 238 5ml
77531 TCEP-HCI,0.5MSolution,pH6.6±0.1 5ml 244 50ml
77534 Glycogen,20mg/ml,Ultrapure 1ml 220 5x1ml
78020Y RNaseA,Lyophilized 100mg 79,163
78040Y E. coliRNAPolymeraseHoloenzyme 100units 91,159
78250 ExoSAP-ITPCRProductCleanup 20reactions 49,14878200 100reactions78201 500reactions78202 2,000reactions78205 5,000reactions
78260 SBECleanupReagent 864µl 52
78300 T4EndonucleaseVII 50,000units 75
78303Y TopoisomeraseII,Alpha 200units 9578303Z 500units
78306 M-MLVReverseTranscriptase 25,000units 90,162 100,000units
78310 Uracil-DNAGlycosylase,Heat-Labile 100units 67
78314 ShrimpDNase,Recombinant 100units 74 500units
78331 10XDNaseIBuffer 1ml 74
78334X OptiKinase 500units 6878334Y 1,000units78334Z 2,500units
78350 One-StepRT-PCRKit 50reactions 47
Code Product Size Page Code Product Size Page
Product Code Index | Numerical
265For bulk or alternate pack sizes, email us at [email protected].
78355 Two-StepRT-PCRKit 50RTrctnsand 48 100PCRrctns
78370 RubyTaqDNAPolymerase 100reactions 45 250units 1,000units 5,000units
78390 ShrimpAlkalinePhosphatase(SAP) 500units 83 1,000units 5,000units
78395 HTExoSAP-ITHigh-ThroughputPCR 480rctnsx8-tubestrip 50,149 ProductCleanup 5,760rctnsx1plate (12x8-tubestrips) 23,040rctns-4plates 1,000rctns(2ml) 5,000rctns(10ml)
78400 Ligate-ITRapidLigationKit 25reactions 12578410 100reactions
78411 rDNaseI,RNase-Free 1,000units 73 2,500units
78430 SequenaseRandomPrimerLabelingKit 30reactions 127
78450 Pyrophosphatase,Inorganic(Recombinant)(rPPase) 5units 85 50units
78454 HumanApurinic/ApyrimidinicEndonuclease1 1,000units 75 (APE1) 5,000units
78480 Change-ITMultipleMutationSiteDirected 20reactions 126 MutagenesisKit
78500 ThermoSequenaseCycleSequencingKit 100reactions 172
78641 MagnesiumChloride(MgCl2),1MSolution, 10x1ml 226 Ultrapure 100ml
78706 PrepEaseMidiPlasmidKit 100preps 140
78711 PrepEaseMaxiPlasmidKit 50preps 140
78720 PrepEaseGigaPlasmidKit 5preps 140
78722 PrepEaseBACPurificationKit 10preps 145
78730 PrepEaseEndotoxin-FreeMaxiPlasmidKit 10preps 142
78737 PrepEaseMiniSpinPlasmidKit 250preps 141
78742 PrepEaseQuickMiniSpinPlasmidKit 250preps 141
78745 PrepEasePlasmid8-wellStripKit 12strips 143
78751 PrepEasePlasmid96-wellPlateKit 4x96-wellplates 144
78753 PrepEasePlasmid96-wellPlateCoreKit 24x96-wellplates 144
78756 PrepEaseGelExtractionKits 50preps 14678757 250preps
78758 PrepEaseDNACleanupKits 50preps 14678759 250preps
78761 PrepEasePCRPurification96-WellPlateKits 10x96-wellplates 14878762 (Ultrafiltration) 50x96-wellplates
78766 PrepEaseRNASpinKits 50preps 15078767 250preps
78776 PrepEaseSequencingDyeCleanupKits 50preps 15278777 250preps
78780 PrepEaseVacuumManifold 1ea 143
78784 PrepEase8-wellStripStarterKitforVacuum 1pack 143
78791 PrepEaseHistidine-TaggedProteinPurificationMini 50preps 153 Kit-HighSpecificity
78793 PrepEaseHistidine-TaggedProteinPurificationMidi 50preps 153 Kit-HighSpecificity
78795 PrepEaseHistidine-TaggedProteinPurificationMaxi 25preps 153 Kit-HighSpecificity
78796 PrepEaseHistidine-TaggedHighSpecificity 30gm 155 PurificationResin
78801 PrepEaseHistidine-TaggedProteinPurificationMini 50preps 154 Kit-HighYield
78803 PrepEaseHistidine-TaggedProteinPurificationMidi 50preps 154 Kit-HighYield
78805 PrepEaseHistidine-TaggedProteinPurificationMaxi 25preps 154 Kit-HighYield
78806 PrepEaseHistidine-TaggedHighYieldPurification 30gm 155 Resin 120gm
78820 PrepEaseProteinPurificationGlutathioneAgarose4B 10ml 155 100ml
78850 PrepEaseGenomicDNAIsolationKits 50preps 14778855 250preps
78871 PrepEaseRNA/ProteinSpinKit 50preps 151
78878 PrepEasemRNAMiniSpinKit 12preps 150
79010 Biotin-11-ψTPAnalog(RNALabeling 250nmol(10mM) 57 Reagent,RLR) 1,000nmol(10mM)
79015 Biotin-11-dXTPAnalog(DNALabeling) 250nmol(10mM) 57 Reagent,DLR 1,000nmol(10mM)
79220 PrepEaseYeastPlasmidIsolationKit 50preps 145
79260 ThermoSequenaseDyePrimerManualCycle 50reactions 173 SequencingKit
798001KT Prep2SeqMultiplexOligoAdaptersforIllumina Set1124,171 platform,(Containsadaptersequences1-12)
798002KT Prep2SeqMultiplexOligoAdaptersforIllumina Set2124,171 platform,(Containsadaptersequences13-27)
79900 Prep2SeqDNALibraryPrepKitfor 10reactions123,170 Illuminaplatform 20reactions
AY1000 EMSAKitwithoutProbeSet kit 103
AY1XXX* EMSAKit-*Seepage104forfulllisting kit 103
AY1XXXP* EMSATFProbeSets-*Seepage104forfulllisting ea 103
AY2002 NuclearExtractionKit 20rxnfromcellculture; 102 10rxnfromwholetissue
AY9999 EMSAComboKit(anythreeprobes) kit 103
BC1009 AFPELISAKit kit 134
BC1013 CA125ELISAKit kit 134
BC1015 CA15-3ELISAKit kit 134
BC1017 CA19-9ELISAKit kit 134
CI0002 CancerCellIsolationKit(2) 2assays 101
Code Product Size Page Code Product Size Page
Product Code Index | Numerical
266 888-362-2447 | 216-765-5000 | usb.affymetrix.com
CI0004 CancerCellIsolationKit(4) 4assays 101
CI0010 CancerCellIsolationKit(10) 10assays 101
DX0001 DeliverXsiRNATransfectionEvaluationKit 0.12ml 98
DX0002 DeliverXsiRNATransfectionKit 0.4ml 98
DX0003 DeliverXsiRNATransfectionKit 1.0ml 98
DX0004 DeliverXsiRNATransfectionKit 4x1.0ml 98
DX0051 DeliverXPlussiRNATransfectionEvaluationKit 0.12ml 98
DX0052 DeliverXPlussiRNATransfectionKit 0.4ml 98
DX0053 DeliverXPlussiRNATransfectionKit 1.0ml 98
DX0054 DeliverXPlussiRNATransfectionKit 4x1.0ml 98
DX0100 FAM-labeledsiRNAControl 0.12ml 98
DX0101 HumanGAPDHsiRNAControl 0.06ml 98
DX0500 DeliverXsiRNABuffer-1 4ml 98
DX0501 DeliverXsiRNABuffer-1 14ml 98
DX0502 DeliverXsiRNABuffer-1 34ml 98
DX0503 DeliverXsiRNABuffer-2 4ml 98
DX0504 DeliverXsiRNABuffer-2 14ml 98
DX0505 DeliverXsiRNABuffer-2 34ml 98
DX0551 DeliverXPlussiRNABuffer-1 4ml 98
DX0552 DeliverXPlussiRNABuffer-1 14ml 98
DX0553 DeliverXPlussiRNABuffer-1 34ml 98
DX0554 DeliverXPlussiRNABuffer-2 4ml 98
DX0555 DeliverXPlussiRNABuffer-2 14ml 98
DX0556 DeliverXPlussiRNABuffer-2 34ml 98
DX1001 DeliverXPeptideTransfectionEvaluationKit 0.6ml 99
DX1002 DeliverXPeptideTransfectionKit 3.0ml 99
DX1100 DeliverXTAMRA-labeledPeptideControl 75µl 99
DX1101 DeliverXPre-formedPeptideTransfectionComplexControl 1.5ml 99
DX1500 DeliverXPeptideBuffer-1 4ml 99
DX1501 DeliverXPeptideBuffer-1 20ml 99
DX1502 DeliverXPeptideBuffer-2 4ml 99
DX1503 DeliverXPeptideBuffer-2 20ml 99
EK0500 HIFColdControlProbe 25μl 110
EK0501 HIFControlCos1NuclearExtract 20μl 110
EK0502 EREColdProbe 25μl 110
EK0503 ERControlMCF7NuclearExtract 20μl 110
EK0504 AP-1ColdProbe 25μl 110
EK0505 ControlHEK292PMANuclearExtract 20μl 110
EK0506 p53ColdControlProbe 25μl 110
EK0508 GRColdControlProbe 25μl 110
EK0509 GRControlHela(DEX)NuclearExtract 20μl 110
EK0510 STAT-1ColdControlProbe 25μl 110
EK0511 STAT-1ControlMCF7(IFNg) 20μl 110
EK0512 MEF-2ColdControlProbe 25μl 110
EK0513 MEF-2ControlNuclearExtract 20μl 110
EK0514 SP-1ColdProbe 25μl 110
EK0515 SP-1ControlRecombinantProtein 20μl 110
EK0516 PPARColdControlProbe 25μl 110
EK0518 PPARaColdControlProbe 25μl 110
EK0519 PPARaControlCos1NuclearExtract 20μl 110
EK0520 GATA-1ColdProbe 25μl 110
EK0521 GATA-1ControlRecombinantProtein 20μl 110
EK0522 NFkBColdControlProbe 25μl 110
EK0523 NFkBControlNuclearExtract 20μl 110
EK1020 HIF-1aELISAKit kit 110
EK1021 HIF-1aELISAKitwithNuclearExtractionKit kit 110
EK1030 ERaELISAKit kit 110
EK1031 ERaELISAKitwithNuclearExtractionKit kit 110
EK1040 AP-1ELISAKit kit 110
EK1041 AP-1ELISAKitwithNuclearExtractionKit kit 110
EK1050 p53ELISAKit kit 110
EK1051 p53ELISAKitwithNuclearExtractionKit kit 112
EK1060 GRELISAKit kit 110
EK1061 GRELISAKitwithNuclearExtractionKit kit 110
EK1070 STAT-1ELISAKit kit 110
EK1071 STAT-1ELISAKitwithNuclearExtractionKit kit 110
EK1080 MEF-2ELISAKit kit 110
EK1081 MEF-2ELISAKitwithNuclearExtractionKit kit 110
EK1090 SP-1ELISAKit kit 110
EK1091 SP-1ELISAKitwithNuclearExtractionKit kit 110
EK1100 GATA-1ELISAKit kit 110
EK1101 GATA-1ELISAKitwithNuclearExtractionKit kit 110
EK1110 NFkBp50ELISAKit kit 110
EK1111 NFkBp50ELISAKitwithNuclearExtractionKit kit 110
EK1120 NFkBp65ELISAKit kit 110
EK1121 NFkBp65ELISAKitwithNuclearExtractionKit kit 110
EK1310 PPARaELISAKit kit 110
EK1311 PPARaELISAKitwithNuclearExtractionKit kit 110
Code Product Size Page Code Product Size Page
Product Code Index | Numerical
267For bulk or alternate pack sizes, email us at [email protected].
EK1400 FKHRELISAKit kit 110
EK1401 FKHRELISAKitwithNuclearExtractionKit kit 110
EK1500 SMAD-4ELISAKit kit 110
EK1501 SMAD-4ELISAKitwithNuclearExtractionKit kit 110
EK1600 SMAD-2ELISAKit kit 110
EK1601 SMAD-2ELISAKitwithNuclearExtractionKit kit 110
EK1800 HNF-3bELISAKit kit 110
EK1801 HNF-3bELISAKitwithNuclearExtractionKit kit 110
EK2000 E2F-1ELISAKit kit 110
EK2001 E2F-1ELISAKitwithNuclearExtractionKit kit 110
EK3100 NFkBp52ELISAKit kit 110
EK3101 NFkBp52ELISAKitwithNuclearExtractionKit kit 110
EK4100 NFkBc-RelELISAKit kit 110
EK4101 NFkBc-RelELISAKitwithNuclearExtractionKit kit 110
EK5100 NFkBRel-bELISAKit kit 110
EK5101 NFkBRel-bELISAKitwithNuclearExtractionKit kit 110
LR0000 TA-luc(EmptyControl)LuciferaseReporterVector 20µg 112
LR0002 AP1(1)LuciferaseReporterVector 10µg 112
LR0003 AP1(2)LuciferaseReporterVector 10µg 112
LR0005 AP3LuciferaseReporterVector 10µg 112
LR0007 ARLuciferaseReporterVector 10µg 112
LR0016 E2F(1)LuciferaseReporterVector 10µg 112
LR0017 E2F(2)LuciferaseReporterVector 10µg 112
LR0019 ELK1LuciferaseReporterVector 10µg 112
LR0020 ERLuciferaseReporterVector 10µg 112
LR0025 GASLuciferaseReporterVector 10µg 112
LR0026 GAS/ISRELuciferaseReporterVector 10µg 112
LR0027 GATALuciferaseReporterVector 10µg 112
LR0028 GATA1LuciferaseReporterVector 10µg 112
LR0029 GATA1/GATA2LuciferaseReporterVector 10µg 112
LR0030 GATA2LuciferaseReporterVector 10µg 112
LR0031 GATA3LuciferaseReporterVector 10µg 112
LR0032 GATA4LuciferaseReporterVector 10µg 112
LR0033 GRLuciferaseReporterVector 10µg 112
LR0038 HSELuciferaseReporterVector 10µg 112
LR0039 IRF1LuciferaseReporterVector 10µg 112
LR0040 ISRELuciferaseReporterVector 10µg 112
LR0043 MEF1LuciferaseReporterVector 10µg 112
LR0044 MEF2LuciferaseReporterVector 10µg 112
LR0045 MEF3LuciferaseReporterVector 10µg 112
LR0050 NF-ATLuciferaseReporterVector 10µg 112
LR0051 NFkB(1)LuciferaseReporterVector 10µg 112
LR0052 NFkB(2)LuciferaseReporterVector 10µg 112
LR0057 p53LuciferaseReporterVector 10µg 112
LR0067 PRLuciferaseReporterVector 10µg 112
LR0068 RARLuciferaseReporterVector 10µg 112
LR0070 RXRLuciferaseReporterVector 10µg 112
LR0071 Sp1LuciferaseReporterVector 10µg 112
LR0072 SRELuciferaseReporterVector 10µg 112
LR0073 SRFLuciferaseReporterVector 10µg 112
LR0075 STAT1p84/p91LuciferaseReporterVector 10µg 112
LR0077 STAT3(1)LuciferaseReporterVector 10µg 112
LR0079 STAT4LuciferaseReporterVector 10µg 112
LR0087 VDRLuciferaseReporterVector 10µg 112
LR0090 YY1LuciferaseReporterVector 10µg 112
LR0093 CRE(1)LuciferaseReporterVector 10µg 112
LR0094 E2F1(3)LuciferaseReporterVector 10µg 112
LR0114 RXR(2)LuciferaseReporterVector 10µg 112
LR0115 Smad3/Smad4LuciferaseReporterVector 10µg 112
LR0116 SmadLuciferaseReporterVector 10µg 112
LR0117 SP1(2)LuciferaseReporterVector 10µg 112
LR0127 STAT1(2)LuciferaseReporterVector 10µg 112
LR0128 HIF1LuciferaseReporterVector 10µg 112
LR1001 ABL1GenePromoterReporterVector 10µg 111
LR1002 BRCA1GenePromoterReporterVector 10µg 111
LR1003 CGPR(CALCR)GenePromoterReporterVector 10µg 111
LR1004 GAGE1GenePromoterReporterVector 10µg 111
LR1005 IRF7GenePromoterReporterVector 10µg 111
LR1006 POMCGenePromoterReporterVector 10µg 111
LR1007 ATF2GenePromoterReporterVector 10µg 111
LR1008 STAT1GenePromoterReporterVector 10µg 111
LR1009 14-3-3-sigma(SFN)GenePromoterReporterVector 10µg 111
LR1010 CIITAGenePromoterReporterVector 10µg 111
LR1011 IFNgGenePromoterReporterVector 10µg 111
LR1012 IGRPGenePromoterReporterVector 10µg 111
LR1013 IL1aGenePromoterReporterVector 10µg 111
LR1014 IL2GenePromoterReporterVector 10µg 111
LR1015 TNFaGenePromoterReporterVector 10µg 111
Code Product Size Page Code Product Size Page
Product Code Index | Numerical
268 888-362-2447 | 216-765-5000 | usb.affymetrix.com
LR1016 SURVIVINGenePromoterReporterVector 10µg 111
LR1017 ERaGenePromoterReporterVector 10µg 111
LR1018 RB1GenePromoterReporterVector 10µg 111
LR1019 APCGenePromoterReporterVector 10µg 111
LR1020 CFTRGenePromoterReporterVector 10µg 111
LR1021 G6PDGenePromoterReporterVector 10µg 111
LR1022 hMLH1GenePromoterReporterVector 10µg 111
LR1023 IL4GenePromoterReporterVector 10µg 111
LR1024 JUNBGenePromoterReporterVector 10µg 111
LR1025 MyoDGenePromoterReporterVector 10µg 111
LR1026 VHLGenePromoterReporterVector 10µg 111
LR1027 v-mycGenePromoterReporterVector 10µg 111
LR1028 MDR1GenePromoterReporterVector 10µg 111
LR1029 COX2(PTGS2)GenePromoterReporterVector 10µg 111
LR1030 HOXA2GenePromoterReporterVector 10µg 111
LR1031 HOXA5GenePromoterReporterVector 10µg 111
LR1032 FHITGenePromoterReporterVector 10µg 111
LR1033 THBS2GenePromoterReporterVector 10µg 111
LR1034 TIMP3GenePromoterReporterVector 10µg 111
MA1210 Protein/DNAArrayI kit 116
MA1211 Protein/DNAArrayII kit 116
MA1212 Protein/DNAArrayIII kit 116
MA1213 Protein/DNAArrayIV kit 116
MA1214 Protein/DNAArrayV kit 116
MA1215 ComboProtein/DNAArray kit 116
MA1310 Protein/DNAArrayIRefillKit kit 116
MA1311 Protein/DNAArrayIIRefillKit kit 116
MA1312 Protein/DNAArrayIIIRefillKit kit 116
MA1313 Protein/DNAArrayIVRefillKit kit 116
MA1314 Protein/DNAArrayVRefillKit kit 116
MA1315 ComboProtein/DNARefillKit kit 116
MA1500 cAMP/CalciumProtein/DNAArray kit 115
MA1505 CellGrowthProtein/DNAArray kit 115
MA1510 NuclearReceptorProtein/DNAArray kit 115
MA1600 cAMP/CalciumProtein/DNAArrayRefill kit 115
MA1605 CellGrowthProtein/DNAArrayRefill kit 115
MA1610 NuclearReceptorProtein/DNAArrayRefill kit 115
MA5010 TF-TFInteractionArrayI kit 115
MA5011 TF-TFInteractionArrayII kit 115
MA5012 TF-TFInteractionArrayIII kit 115
MA5041 PPARa-TFInteractionArrayI kit 118
MA5042 PPARa-TFInteractionArrayII kit 118
MA5043 PPARa-TFInteractionArrayIII kit 118
MA5044 PPARg-TFInteractionArrayI kit 118
MA5045 PPARg-TFInteractionArrayII kit 118
MA5046 PPARg-TFInteractionArrayIII kit 118
MA5051 NFkBp50-TFInteractionArrayI kit 118
MA5052 NFkBp50-TFInteractionArrayII kit 118
MA5053 NFkBp50-TFInteractionArrayIII kit 118
MA5054 NFkBp65-TFInteractionArrayI kit 118
MA5055 NFkBp65-TFInteractionArrayII kit 118
MA5056 NFkBp65-TFInteractionArrayIII kit 118
MA5110 TF-TFInteractionIRefill kit 115
MA5111 TF-TFInteractionIIRefill kit 115
MA5112 TF-TFInteractionIIIRefill kit 115
MA6110 HumanCytokineAntibodyArray1.0 2membranes 134
MA6120 HumanCytokineAntibodyArray1.0 4membranes 134
MA6130 HumanCytokineAntibodyArray1.0 6membranes 134
MA6140 HumanCytokineAntibodyArray1.0 8membranes 134
MA6150 HumanCytokineAntibodyArray3.0 4membranes 134
MA6160 HumanCytokineAntibodyArray3.0 8membranes 134
MA6310 HumanAngiogenesisAntibodyArray kit 133
MA6320 MouseAngiogenesisAntibodyArray kit 133
MA6410 MouseCytokineAntibodyArray1.0 4membranes 134
MA6412 MouseCytokineAntibodyArray1.0 8membranes 134
ME0001 PPARaCMVExpressionVector 15µg 114
ME0002 PPARb(PPARΔ)CMVExpressionVector 15µg 114
ME0003 PPARg-1CMVExpressionVector 15µg 114
ME0004 PPARg-2CMVExpressionVector 15µg 114
ME0005 NFkBp50CMVExpressionVector 15µg 114
ME0006 NFkBp65(RelA)CMVExpressionVector 15µg 114
ME0007 GR(NR3C1)CMVExpressionVector 15µg 114
ME0010 E2F2CMVExpressionVector 15µg 114
ME0014 IRF1CMVExpressionVector 15µg 114
ME0018 MAFKCMVExpressionVector 15µg 114
ME0019 MAXCMVExpressionVector 15µg 114
ME0020 CITED1CMVExpressionVector 15µg 114
ME0022 NF-YCCMVExpressionVector 15µg 114
Code Product Size Page Code Product Size Page
Product Code Index | Numerical
269For bulk or alternate pack sizes, email us at [email protected].
ME0024 NR1I2CMVExpressionVector 15µg 114
ME0025 NR1I3CMVExpressionVector 15µg 114
ME0029 PREBCMVExpressionVector 15µg 114
ME0032 SRYCMVExpressionVector 15µg 114
ME0033 STAT1CMVExpressionVector 15µg 114
ME0035 STAT5BCMVExpressionVector 15µg 114
ME0040 USF1CMVExpressionVector 15µg 114
ME0043 ERaCMVExpressionVector 15µg 114
ME0044 FOSCMVExpressionVector 15µg 114
ME0045 JUNCMVExpressionVector 15µg 114
ME0046 NF-ATC1CMVExpressionVector 15µg 114
ME0047 p53CMVExpressionVector 15µg 114
ME0049 GATA1CMVExpressionVector 15µg 114
ME0052 MYCCMVExpressionVector 15µg 114
ME0054 PBX1CMVExpressionVector 15µg 114
ME0055 ATF3CMVExpressionVector 15µg 114
ME0059 ATF1CMVExpressionVector 15µg 114
ME0060 CDX2CMVExpressionVector 15µg 114
ME0061 NR0B1CMVExpressionVector 15µg 114
ME0062 HNF4gCMVExpressionVector 15µg 114
ME0065 BTG2CMVExpressionVector 15µg 114
ME0066 GFI1CMVExpressionVector 15µg 114
ME0071 SMAD1(MADH1)CMVExpressionVector 15µg 114
MP1100 PromoterMethylationPCRKit kit 24
MP1101 14-3-3 ea 24
MP1102 14-3-3 ea 24
MP1103 14-3-3 ea 24
MP1104 14-3-3 ea 24
MP1105 ABL1 ea 24
MP1106 ABL1 ea 24
MP1107 BAGE ea 24
MP1108 BAGE ea 24
MP1109 BAGE ea 24
MP1110 BAGE ea 24
MP1111 BAGE ea 24
MP1112 BAGE ea 24
MP1113 BRCA1 ea 24
MP1114 BRCA1 ea 24
MP1115 BRCA1 ea 24
MP1116 BRCA1 ea 24
MP1117 CalcitoninCGPR ea 24
MP1118 CalcitoninCGPR ea 24
MP1119 CalcitoninCGPR ea 24
MP1120 CalcitoninCGPR ea 24
MP1121 CD14 ea 24
MP1122 CD14 ea 24
MP1123 CDKN2A ea 24
MP1124 CDKN2A ea 24
MP1125 CDKN2A ea 24
MP1126 CDKN2A ea 24
MP1127 COX2 ea 24
MP1128 COX2 ea 24
MP1129 CyclinD2 ea 24
MP1130 CyclinD2 ea 24
MP1131 CyclinD2 ea 24
MP1132 CyclinD2 ea 24
MP1133 DAPK ea 24
MP1134 DAPK ea 24
MP1135 DAPK ea 24
MP1136 DAPK ea 24
MP1137 glut4 ea 24
MP1138 glut4 ea 24
MP1139 H-ras ea 24
MP1140 H-ras ea 24
MP1141 H-ras ea 24
MP1142 H-ras ea 24
MP1143 INFg ea 24
MP1144 INFg ea 24
MP1145 GAGE1 ea 24
MP1146 GAGE1 ea 24
MP1147 GAGE1 ea 24
MP1148 GAGE1 ea 24
MP1149 HOXA2 ea 25
MP1150 HOXA2 ea 25
MP1151 IL4 ea 25
MP1152 IL4 ea 25
MP1153 IRF7 ea 25
MP1154 IRF7 ea 25
Code Product Size Page Code Product Size Page
Product Code Index | Numerical
270 888-362-2447 | 216-765-5000 | usb.affymetrix.com
MP1155 NIS ea 25
MP1156 NIS ea 25
MP1157 NIS ea 25
MP1158 NIS ea 25
MP1159 NME2 ea 25
MP1160 NME2 ea 25
MP1161 NME2 ea 25
MP1162 NME2 ea 25
MP1163 PDGFB ea 25
MP1164 PDGFB ea 25
MP1165 POMC ea 25
MP1166 POMC ea 25
MP1167 NES ea 25
MP1168 NES ea 25
MP1169 NES ea 25
MP1170 NES ea 25
MP1171 PRA2 ea 25
MP1172 PRA2 ea 25
MP1173 TFF1 ea 25
MP1174 TFF1 ea 25
MP1175 VHL ea 25
MP1176 VHL ea 25
MP1177 Survivin ea 25
MP1178 Survivin ea 25
MP1179 CASP8 ea 25
MP1180 CASP8 ea 25
MP11XX PCRPrimers(forPromoterMethylationPCRKit) ea 24
RC0002 NFkBA549ReporterStableCellLine vial 122
RC0004 STAT-1HeLaReporterStableCellLine vial 122
RC0005 NFATHeLaReporterStableCellLine vial 122
RC0006 AP-1293ReporterStableCellLine vial 122
RC0007 CREB293ReporterStableCellLine vial 122
RC0008 CREBCHOReporterStableCellLine vial 122
RC0009 NFATK562ReporterStableCellLine vial 122
RC0012 SRFHeLaReporterStableCellLine vial 122
RC0013 NFkBHeLaReporterStableCellLine vial 122
RC0014 NFkB293ReporterStableCellLine vial 122
RC0015 NFkBNIH3T3ReporterStableCellLine vial 122
RC0016 NFkBC2C12ReporterStableCellLine vial 122
RC0017 HIFNIH3T3ReporterStableCellLine vial 122
RC0018 GR293ReporterStableCellLine vial 122
RC0019 GRHeLaReporterStableCellLine vial 122
RC1001 TAD-KinaseReporterStableCellLine vial 122
RC2001 CHO/GFP-NFkBp65StableCellLine vial 122
RC2002 CHO/GFP-GRaStableCellLine vial 122
RC5001 NFkBp50KnockdownHeLaStableCellLine vial 122
Product Name Index | Alphabetical
271For bulk or alternate pack sizes, email us at [email protected].
Product Page Product Page
A
14-3-3 24
14-3-3-sigma(SFN)GenePromoterReporterVector 111
ABL1 24
ABL1GenePromoterReporterVector 111
N-Acetyl-L-Cysteine 188
S-AcetylCoenzymeA,LithiumSalt,Ultrapure 188
AcetylatedBSA—SeeAlbumin,Bovine,Acetylated 198
Acrylamide 188
Acrylamide,Ultrapure 188
AcrylamideGelMixesandSolutions
GlycerolTolerantGel(GTG)Buffer,20XSolution 219
RapidGel™40%LiquidAcrylamideStockSolution 237
RapidGel-XL6%DNASequencingGelSolution 238
StopSolution 242
Adenine 188
AdenineSulfate,Dihydrate 188
Adenosine 189
Adenosine-5’-Diphosphate,DisodiumSalt,Dihydrate 189
Adenosine-5’-Monophosphate,DisodiumSalt 189
Adenosine-5’-Monophosphate,FreeAcid 189
Adenosine-5’-Monophosphate,FreeAcid,Monohydrate 189
Adenosine-5’-Triphosphate,100mMSolution,Ultrapure 61,189
Adenosine-5’-Triphosphate,DisodiumSalt 189
S-Adenosyl-L-MethionineSulfatep-Toluenesulfonate 190
Adonitol 190
ADP—SeeAdenosine-5’-Diphosphate,DisodiumSalt,Dihydrate 189
AEBSFHydrochloride 190
AFPELISAKit 134
Agar 190
Agar,Bacteriological,Ultrapure 190
Agar,NobleAgar,Ultrapure 190
Agarose-HE,Ultrapure 192
Agarose-Hi-Res,Separation≤1000bp,Ultrapure 195
Agarose-LE,Ultrapure 194
Agarose-LowMeltSeparation≥1000bp,GeneticPerformanceCertified,Ultrapure 196
Agarose-LowMeltSeparation≤1000bp,GeneticPerformanceCertified,Ultrapure 197
Agarose-ME,Ultrapure 192
Agarose-Separation≥500bp,GeneticPerformanceCertified,Ultrapure 193
Agarose-UltraLowMelt,Ultrapure 197
b-Alanine 198
Albumin,Bovine,Acetylated,Ultrapure 198
Albumin,Bovine,CohnFractionV,pH5.2 198
Albumin,Bovine,Crystallized,pH5.2 198
Albumin,Bovine,pH7 198
Albumin,Bovine,Protease-Free 198
Albumin,Bovine,RIAGrade,pH7 198
Albumin,Egg 199
AlcoholDehydrogenase 199
AlkalinePhosphatases
CalfIntestinalAlkalinePhosphatase(CIAP) 82
ShrimpAlkalinePhosphatase(SAP) 83
SuperSAPShrimpAlkalinePhosphatase 84
4-AminoAntipyrene 199
p-AminoBenzoicAcid(PABA) 199
6-AminoPurine—SeeAdenine 188
L-a-Amino-N-ButyricAcid 199
AminoaceticAcid—SeeGlycine 219
2-AminoethaneSulfonicAcid—SeeTaurine 243
L-a-AminoglutaricAcid—SeeL-GlutamicAcid 218
AmmoniumAcetate,Ultrapure 199
AmmoniumAcetate(NH4OAc),5MSolution,Ultrapure 199
AmmoniumPersulfate(APS),Ultrapure 200
AmmoniumSulfate,Ultrapure 200
AMP—SeeAdenosine-5’-Monophosphate,DisodiumSalt 189
cAMP/CalciumProtein/DNAArray 115
cAMP/CalciumProtein/DNAArrayRefill 115
Ampicillin,SodiumSalt,Ultrapure 200
AMVReverseTranscriptase 90,162
AntibioticG-418Sulfate—SeeG-418Sulfate 217
AP-1293ReporterStableCellLine 122
AP-1ColdProbe 110
AP-1ELISAKit 110
AP-1ELISAKitwithNuclearExtractionKit 110
AP1(1)LuciferaseReporterVector 112
AP1(2)LuciferaseReporterVector 112
AP3LuciferaseReporterVector 112
APCGenePromoterReporterVector 111
Aprotinin,Ultrapure 200
Product Name Index | Alphabetical
272 888-362-2447 | 216-765-5000 | usb.affymetrix.com
Product Page Product Page
ARLuciferaseReporterVector 112
L-Arabinose 200
L-(+)-Arabinose 200
L-Arginine 200
L-Arginine,Hydrochloride 201
L-AscorbicAcid 201
L-Asparagine,Anhydrous 201
L-Asparagine,Monohydrate 201
L-AsparticAcid 201
L-AsparticAcid,MagnesiumSalt,Dihydrate 201
L-AsparticAcid,SodiumSalt,Monohydrate 201
ATF1CMVExpressionVector 114
ATF2GenePromoterReporterVector 111
ATF3CMVExpressionVector 114
ATP—SeeAdenosine-5’-Triphosphate 189
Avidin-AlkalinePhosphataseConjugate—SeeStreptavidin-AlkalinePhosphataseConjugate 242
B
Bacitracin 202
BacterialCultureMedia
LBAgar,Ready-MadePowder 224
LBBroth,Ready-MadePowder 224
LuriaAgar,Ready-MadePowder 225
LuriaBroth,Ready-MadePowder 225
NZCYMBroth,Ready-MadePowder 231
TerrificBroth,Ready-MadePowder 245
2XYTBroth 251
BacteriologicalPeptone,Ultrapure 202
BAGE 24
BCIP—See5-Bromo-4-Chloro-3-IndolylPhosphate 203
Betaine,5MSolution,Ultrapure 202
Bicine 202
BindingProteins
RecAProtein 65
Single-StrandedDNABindingProtein(SSB) 65
T4Gene32Protein 66
Biotin-11-dXTPAnalog(DNALabelingReagent,DLR) 57
Biotin-11-ψTPAnalog(RNALabelingReagent,RLR) 57
Bis-Acrylamide—SeeN,N’-Methylene-bis-Acrylamide 228
Bis-Tris,Ultrapure 202
BlottingReagents
Denhardt’sSolution,50X 209
SSC,20XSolution 241
SSPE,20XSolution 241
TBS,20XSolution,pH7.4 244
TBSwithTween(TBST),20XSolution,pH7.4 244
BoricAcid,Ultrapure 202
BRCA1 24
BRCA1GenePromoterReporterVector 111
BrewersYeast—SeeYeast,Brewers 251
BrilliantBlueG,Ultrapure 203
BrilliantBlueR,Ultrapure 203
5-Bromo-4-Chloro-3-5-Bromo-4-Chloro-3-IndolylPhosphate,DisodiumSalt,Ultrapure 203
5-Bromo-4-Chloro-3-Indolyl-a-D-Galactopyranoside,Ultrapure 203
5-Bromo-4-Chloro-3-Indolyl-b-D-Galactopyranoside,Ultrapure 203
5-Bromo-4-Chloro-3-IndolylBromocresolGreen,SodiumSalt,Ultrapure 203
BromophenolBlue,SodiumSalt,Ultrapure 204
BSA—SeeAlbumin,Bovine 198
BTG2CMVExpressionVector 114
Buffers
GlycerolTolerantGel(GTG)Buffer,20XSolution 219
HEPES,1MSolution,pH7.3 222
PBS,10XSolution,pH7.4 232
RapidRunAgaroseBuffer,20XSolution 238
TAEBuffer,10XSolution 243
TAEBuffer,50XSolution 243
TBEBuffer,5XSolution 243
TBEBuffer,10XReady-MixedPowder 243
TEBuffer,1XSolution 244
TEBuffer,1XSolution,pH7.0 244
TEBuffer,1XSolution,pH7.0,LowEDTA 244
TEBuffer,1XSolution,pH8.0 244
TEBuffer,1XSolution,pH8.0,LowEDTA 244
TEBuffer,50XSolution 244
Tris-Glycine(TG)Buffer,10XSolution 247
Tris-Glycine-SDS(TGS)Buffer,10XSolution 247
Tris/HCl,1MSolution,pH7.0 248
Tris/HCl,1MSolution,pH7.5 248
Tris/HCl,1MSolution,pH8.0 248
Product Name Index | Alphabetical
273For bulk or alternate pack sizes, email us at [email protected].
Product Page Product Page
C
CA15-3ELISAKit 134
CA19-9ELISAKit 134
CA125ELISAKit 134
CacodylicAcid,SodiumSaltTrihydrate 204
Caffeine,Anhydrous,Ultrapure 204
CalciumChloride,Dihydrate 204
D-CalciumPantothenate 204
CalcitoninCGPR 24
CalfIntestinalAlkalinePhosphatase(CIAP) 82
CancerCellIsolationKit(2) 101
CancerCellIsolationKit(4) 101
CancerCellIsolationKit(10) 101
Carbenicillin,DisodiumSalt 205
CarboxypeptidaseY 205
Casein,Hammarsten,Ultrapure 205
Casein(HighNitrogen) 205
CaseinHydrolysate,EnzymaticDigest,Ultrapure 205
Casein,Sodium 205
Casein-Vitafree,Ultrapure 205
CASP8 25
Catalase 205
CD14 24
CDKN2A 24
CDX2CMVExpressionVector 114
CellGrowthProtein/DNAArray 115
CellGrowthProtein/DNAArrayRefill 115
CellularManipulation
DeliverXPeptideTransfectionKits 99
CancerCellIsolationKit 101
Cellulase 205
CesiumChloride,Ultrapure 206
CetylTrimethylAmmoniumBromide 206
CFTRGenePromoterReporterVector 111
CGPR(CALCR)GenePromoterReporterVector 111
Change-ITMultipleMutationSiteDirectedMutagenesisKit 126
ChickenIntestineAlkalinePhosphastase 206
Chloramphenicol,Ultrapure 206
CHO/GFP-GRaStableCellLine 122
CHO/GFP-NFkBp65StableCellLine 122
Cholesterol 206
CholineChloride 207
CIITAGenePromoterReporterVector 111
CITED1CMVExpressionVector 114
CitricAcid,Anhydrous 207
CitricAcid,TrisodiumSalt,Dihydrate,Ultrapure 207
Cloning
Ligate-ITKit 125
Cocarboxylase 207
CoenzymeI—Seeb-NicotinamideAdenineDinucleotide 229
CoenzymeII—SeeNicotinamideAdenineDinucleotidePhosphate 230
CoenzymeA,FreeAcid,Trihydrate 207
CoenzymeA,TrilithiumSalt,Dihydrate 207
Collagen 207
Collagenase,TypeI 207
ComboProtein/DNAArray 116
ComboProtein/DNARefillKit 116
CompactReactionColumns 207-208
ControlHEK292PMANuclearExtract 110
CoomassieStains—SeeBrilliantBlueG&R 203
COX2 24
COX2(PTGS2)GenePromoterReporterVector 111
CRCLowerFilters 208
CRCUpperFilters 208
CRE(1)LuciferaseReporterVector 112
CreatineKinase 208
CreatinePhosphate,DisodiumSalt,Tetrahydrate 208
CREB293ReporterStableCellLine 122
CREBCHOReporterStableCellLine 122
CTAB—SeeCetylTrimethylAmmoniumBromide 206
CTP—SeeCytidine-5’-Triphosphate 209
Cyanocobalamin—SeeVitaminB12 250
CyclinD2 24
a-Cyclodextrin 208
L-Cystine 208
L-Cysteine,Hydrochloride,Monohydrate 208
Cytidine,FreeBase 208
Cytidine-5’-Triphosphate,DisodiumSalt,Dihydrate 209
CytochromeC 209
Product Name Index | Alphabetical
274 888-362-2447 | 216-765-5000 | usb.affymetrix.com
Product Page Product Page
D
DAPI 209
DAPK 24
7-deaza-2’-Deoxyguanosine-5’-Triphosphate,LithiumSalt,10mMSolution 60
DeliverXsiRNABuffer-1 98
DeliverXsiRNABuffer-2 98
DeliverXsiRNATransfectionEvaluationKit 98
DeliverXsiRNATransfectionKit 98
DeliverXPeptideBuffer-1 99
DeliverXPeptideBuffer-2 99
DeliverXPeptideTransfectionEvaluationKit 99
DeliverXPeptideTransfectionKit 99
DeliverXPlussiRNABuffer-1 98
DeliverXPlussiRNABuffer-2 98
DeliverXPlussiRNATransfectionEvaluationKit 98
DeliverXPlussiRNATransfectionKit 98
DeliverXPre-formedPeptideTransfectionComplexControl 99
DeliverXTAMRA-labeledPeptideControl 99
Denhardt’sSolution,50X,Ultrapure 209
2’-Deoxyadenosine 209
2’-Deoxyadenosine-5’-Triphosphate,SodiumSalt,100mMSolution,Ultrapure 59,210
DeoxycholicAcid,SodiumSalt 210
2’-Deoxycytidine,Hydrochloride,Synthetic 210
2’-Deoxycytidine-5’-Triphosphate,SodiumSalt,100mMSolution,Ultrapure 59,210
2’-Deoxyguanosine 210
2’-Deoxyguanosine,MonohydrateSynthetic 210
7-deaza-2’-Deoxyguanosine-5’-Triphosphate,LithiumSalt,10mMSolution 210
2’-Deoxyguanosine-5’-Triphosphate,SodiumSalt,100mMSolution,Ultrapure 59,210
2’-Deoxynucleoside-5’-TriphosphateMixes—SeePCRNucleotideMixes 232
2’-Deoxynucleoside-5’-Triphosphates,SodiumSalt,100mMSolutions, 59,211SetsofFour,Ultrapure,4x25µmol(250µl)eachdNTPperpack
2’-Deoxynucleoside-5’-Triphosphates,SodiumSalt,100mMSolutions, 59,211SetsofFour,Ultrapure,4x100µmol(1ml)eachdNTPperpack
2’-Deoxynucleoside-5’-Triphosphates,SodiumSalt,100mMSolutions, 59,211SetsofFour,Ultrapure,4x500µmol(5ml)eachdNTPperpack
2-Deoxy-D-Ribose,Ultrapure 211
2’-Deoxythymidine-5’-Triphosphate,SodiumSalt,100mMSolution,Ultrapure 59,211
2’-Deoxyuridine-5’-Triphosphate,SodiumSalt,100mMSolution,Ultrapure 59,211
DEPC—SeeDiethylPyrocarbonate 212
Dextran,Ultrapure 211
DextranSulfate,SodiumSalt 211
DextranSulfate,SodiumSalt,Ultrapure 211
D-(+)-Dextrose,Anhydrous 211
D-(+)-Dextrose,Monohydrate 212
Diaphorase 212
2’,3’-Dideoxyadenosine-5’-Triphosphate,Ultrapure,10mMaqueoussolution, 60,212pH7.5
2’,3’-Dideoxycytidine-5’-Triphosphate,Ultrapure,10mMaqueoussolution, 60,212pH7.5
2’,3’-Dideoxynucleoside-5’-Triphosphates,10mMSolutions,SetsofFour, 60,212Ultrapure,4x0.5µmol(50µl)eachddNTPperpack
2’,3’-Dideoxynucleoside-5’-Triphosphates,10mMSolutions,SetsofFour, 60,212Ultrapure,4x1.0µmol(100µl)eachddNTPperpack
2’,3’-Dideoxynucleoside-5’-Triphosphates,100mMSolutions,SetofFour, 60,212Ultrapure,4x4µmol(40µl)eachddNTPperpack
2’,3’-Dideoxythymidine-5’-Triphosphate,Ultrapure,10mMaqueoussolution, 60,212pH7.5
DiethylPyrocarbonate,Ultrapure 212
DihydrostreptomycinSulfate 213
Dimedone 213
p-DimethylaminoBenzaldehyde 213
p-DimethylaminoCinnamaldehyde 213
Dithioerythritol,Ultrapure 213
Dithiothreitol 213
Dithiothreitol,Ultrapure 213
Dithiothreitol(DTT),0.1MSolution,Ultrapure 213
DNA
DNA,CalfThymus 128
DNA,E. coli 128
DNA,FishSperm 128
DNA,FishSperm,Activated 128
DNA,FishSperm,SodiumSalt 128
DNA-Cellulose,Double-Stranded 128
DNA-Cellulose,Single-Stranded 129
M13mp18,Single-Stranded,0.2µg/ul 129
M13mp18DNA,Single-Stranded,1.0µg/µl 129
Oligo(dT)12-18Primer,MWa4500 129
Oligop(dT)12-18SodiumSalt 129
pUC19DNA,0.2µg/µl 129
pUC19DNA,1.0µg/µl 129
DNACleanup
PrepEaseDNACleanupKits 146
DNALadder,1kbPlus 131
DNALadder,100bp 130
Product Name Index | Alphabetical
275For bulk or alternate pack sizes, email us at [email protected].
Product Page Product Page
DNALoadingBuffer(OXG),6X 130
DNALoadingBuffer(BXF),6X 131
DNAMarkers
DNALadder,1kbPlus 131
DNALadder,100bp 130
PCRMarkers,50-2000bp 130
DNAPolymeraseI 87
DNAPolymerases
DNAPolymeraseI 87
Exonuclease-FreeKlenow 86
Exonuclease-FreeKlenowTrisBuffer 86
KlenowDNAPolymeraseI 87
SequenaseVersion2.0DNAPolymerase 88,174
T7DNAPolymerase 88
TthDNAPolymerase 89
DNA-Cellulose,Double-Stranded 124
DNA-Cellulose,Single-Stranded 124
DNaseI 213
DNaseI,Lyophilized 72
DNaseI,Solution 72
rDNaseI,RNase-Free 73
10XDNaseIBuffer 74
DNases
10XDNaseIBuffer 74
DNaseI,Lyophilized 72
DNaseI,Solution 72
rDNaseI,RNase-Free 73
ShrimpDNase,Recombinant 74
DodecylSulfate,LithiumSalt—SeeLithiumDodecylSulfate 225
DodecylSulfate,SodiumSalt—SeeSodiumDodecylSulfate 240
N-DodecylTrimethylammoniumBromide 214
DPN—SeeNicotinamideAdenineDinucleotide 229
DTAB—SeeN-DodecylTrimethylammoniumBromide 214
DTE—SeeDithioerythritol 213
DTT—SeeDithiothreitol 213
E
E2F-1ELISAKit 110
E2F-1ELISAKitwithNuclearExtractionKit 110
E2F(1)LuciferaseReporterVector 112
E2F2CMVExpressionVector 114
E2F(2)LuciferaseReporterVector 112
E2F1(3)LuciferaseReporterVector 112
E. coliDNALigase 70
E. coliRNAPolymeraseHoloenzyme 91,159
EDTA—SeeEthylenediamineTetraaceticAcid 215
EggWhite—SeeAlbumin-Egg 199
EGTA—SeeEthyleneGlycol-o-o’-bis(2-aminoethyl)-N,N,N’,N’,-TetraaceticAcid 215
Elastase 214
ELK1LuciferaseReporterVector 112
EMSAComboKit(anythreeprobes) 103
EMSAKit-Seepage104forfulllisting 103
EMSAKitwithoutProbeSet 103
EMSATFProbeSets-Seepage104forfulllisting 103
EndPointPCR
RubyTaqDNAPolymerase 13
RubyTaqPCRMasterMix(2X) 14
TaqDNAPolymerase 11
TaqPCRKit 12
TaqPCRMasterMix(2X) 12
Endonucleases
HumanApurinic/ApyrimidinicEndonuclease1(APE1) 75
T4EndonucleaseVII 75
Enolase 214
ERControlMCF7NuclearExtract 110
ERLuciferaseReporterVector 112
ERaCMVExpressionVector 114
ERaELISAKit 110
ERaELISAKitwithNuclearExtractionKit 110
ERaGenePromoterReporterVector 111
EREColdProbe 110
Ergocalciferol 214
Esculin 214
EthidiumBromide,Ultrapure 214
EthidiumBromideDrops,Ultrapure 214
EthylenediaminetetraaceticAcid,(EDTA),0.5MSolution,Ultrapure 215
EthylenediaminetetraaceticAcid,(EDTA)DisodiumSalt,Dihydrate 215
EthylenediaminetetraaceticAcid,(EDTA)DisodiumSalt,Dihydrate,Ultrapure 215
EthyleneDiamineTetraaceticAcid,(EDTA)TetrasodiumSalt,Dihydrate,Ultrapure 215
EthyleneGlycol-O,O’-Bis(2Amino-ethyl)-N,N,N’,N’,-TetraaceticAcid(EGTA),Ultrapure 215
Product Name Index | Alphabetical
276 888-362-2447 | 216-765-5000 | usb.affymetrix.com
Product Page Product Page
ExonucleaseI,Highconcentration,20units/µl 76
ExonucleaseI,Standardconcentration,10units/µl 76
ExonucleaseIII 77
ExonucleaseVII 77
Exonuclease-FreeKlenow 86
Exonucleases
ExonucleaseI 76
ExonucleaseIII 77
ExonucleaseVII 77
T7Gene6Exonuclease 78
ExoSAP-ITPCRProductCleanup 49,148
F
FAM-labeledsiRNAControl 98
FetalBovineSerum(FBS) 215
FHITGenePromoterReporterVector 111
FideliTaqDNAPolymerase 15
FideliTaqPCRMasterMix(2X) 16
FideliTaqRT-PCRMasterMix(2X) 46
First-StrandcDNASynthesisKitforReal-TimePCR 39
FKHRELISAKit 110
FKHRELISAKitwithNuclearExtractionKit 110
FlavinAdenineDinucleotide,DisodiumSalt,Hydrate 216
FluoresceinPassiveReferenceDye 53
FolicAcid 216
Formamide,Ultrapure 216
FOSCMVExpressionVector 114
b-D-Fructose 216
D-Fructose-6-Phosphate,DisodiumSalt 216
G
G-418Sulfate,Ultrapure 217
G6PDGenePromoterReporterVector 111
GAGE1 24
GAGE1GenePromoterReporterVector 111
D-(+)-Galactose 217
GAS/ISRELuciferaseReporterVector 112
GASLuciferaseReporterVector 112
GATALuciferaseReporterVector 112
GATA-1ColdProbe 110
GATA-1ControlRecombinantProtein 110
GATA-1ELISAKit 110
GATA-1ELISAKitwithNuclearExtractionKit 110
GATA1CMVExpressionVector 114
GATA1LuciferaseReporterVector 112
GATA1/GATA2LuciferaseReporterVector 112
GATA2LuciferaseReporterVector 112
GATA3LuciferaseReporterVector 112
GATA4LuciferaseReporterVector 112
GDP—SeeGuanosine-5’-Diphosphate 221
GelElectrophoresisReagents
EthidiumBromideDrops 214
GlycerolTolerantGel(GTG)Buffer,20XSolution 219
RapidRunAgaroseBuffer,20XSolution 238
TAEBuffer,10XSolution 243
TAEBuffer,50XSolution 243
TBEBuffer,5XSolution 243
TBEBuffer,10XReady-MixedPowder 243
Tris-Glycine(TG)Buffer,10XSolution 247
Tris-Glycine-SDS(TGS)Buffer,10XSolution 247
GelMixesandSolutions
GlycerolTolerantGel(GTG)Buffer,20XSolution 219
RapidGel40%LiquidAcrylamideStockSolution 237
RapidGel-XL6%DNASequencingGelSolution 238
StopSolution 242
GelExtraction
PrepEaseGelExtractionKits 146
GenePromoterReporterVectors
14-3-3-sigma(SFN)GenePromoterReporterVector 111
ABL1GenePromoterReporterVector 111
APCGenePromoterReporterVector 111
ATF2GenePromoterReporterVector 111
BRCA1GenePromoterReporterVector 111
CFTRGenePromoterReporterVector 111
CGPR(CALCR)GenePromoterReporterVector 111
CIITAGenePromoterReporterVector 111
COX2(PTGS2)GenePromoterReporterVector 111
ERaGenePromoterReporterVector 111
FHITGenePromoterReporterVector 111
G6PDGenePromoterReporterVector 111
GAGE1GenePromoterReporterVector 111
hMLH1GenePromoterReporterVector 111
Product Name Index | Alphabetical
277For bulk or alternate pack sizes, email us at [email protected].
Product Page Product Page
HOXA2GenePromoterReporterVector 111
HOXA5GenePromoterReporterVector 111
IFNgGenePromoterReporterVector 111
IGRPGenePromoterReporterVector 111
IL1aGenePromoterReporterVector 111
IL2GenePromoterReporterVector 111
IL4GenePromoterReporterVector 111
IRF7GenePromoterReporterVector 111
JUNBGenePromoterReporterVector 111
MDR1GenePromoterReporterVector 111
MyoDGenePromoterReporterVector 111
POMCGenePromoterReporterVector 111
RB1GenePromoterReporterVector 111
STAT1GenePromoterReporterVector 111
SURVIVINGenePromoterReporterVector 111
THBS2GenePromoterReporterVector 111
TIMP3GenePromoterReporterVector 111
TNFaGenePromoterReporterVector 111
VHLGenePromoterReporterVector 111
v-mycGenePromoterReporterVector 111
GeneticinDisulfate—SeeG-418Sulfate 217
GenomicDNAIsolation
PrepEaseGenomicDNAIsolationKit 147
GentamicinSulfate 217
GFI1CMVExpressionVector 114
g-Globulins,FractionII 217
a-D-(+)-Glucose—SeeDextrose 211
GlucoseOxidase 217
D-Glucose-6-Phosphate,DisodiumSalt,Dihydrate 217
Glucose-6-PhosphateDehydrogenase 217,218
Glucose-6-PhosphateGlutamateDehydrogenase 218
glut4 24
L-GlutamicAcid 218
L-GlutamicAcid,MonosodiumSalt,Monohydrate 218
L-Glutamine 218
GlutamylCysteinylGlycine—SeeGlutathione 218
Glutathione-Oxidized 218
Glutathione-Reduced 218
Glycerin—SeeGlycerol 219
Glycerol,Ultrapure 219
GlycerolTolerantGel(GTG)Buffer,20XSolution,Ultrapure 219
b-Glycerophosphate,SodiumSalt,Pentahydrate 219
Glycine 220
Glycine,Ultrapure 219
Glycogen,20mg/ml,Ultrapure 220
Glycogen,Ultrapure 220
Glycosylases
Uracil-DNAGlycosylase,E. coli 67
Uracil-DNAGlycosylase,Heat-Labile 67
Glycyl-Glycine 220
GR293ReporterStableCellLine 122
GRColdControlProbe 110
GRControlHela(DEX)NuclearExtract 110
GRELISAKit 110
GRELISAKitwithNuclearExtractionKit 110
GRHeLaReporterStableCellLine 122
GRLuciferaseReporterVector 112
GR(NR3C1)CMVExpressionVector 114
GST-TaggedProteinPurification
PrepEaseProteinPurificationGlutathioneAgarose4B 155
GTP—SeeGuanosine-5’-Triphosphate 221
GuanidineHydrochloride 221
GuanidineHydrochloride,Ultrapure 220
GuanidineThiocyanate,Ultrapure 221
Guanosine-5’-Diphosphate,DisodiumSalt 221
Guanosine-5’-Triphosphate,DisodiumSalt 221
Guanosine-5’-Triphosphate,SodiumSalt,100mMSolution,Ultrapure 61,221
H
Hemin 221
Heparin,SodiumSalt 221
HEPES,1MSolution,pH7.3,Ultrapure 222
HEPES,SodiumSalt 222
HEPES,Ultrapure 222
Hexokinase 222
HIFColdControlProbe 110
HIFControlCos1NuclearExtract 110
HIFNIH3T3ReporterStableCellLine 122
HIF-1aELISAKit 110
HIF-1aELISAKitwithNuclearExtractionKit 110
Product Name Index | Alphabetical
278 888-362-2447 | 216-765-5000 | usb.affymetrix.com
Product Page Product Page
HIF1LuciferaseReporterVector 112
Histidine-TaggedProteinPurificationKits
PrepEaseHistidine-TaggedMaxiKit–HighSpecificity 153
PrepEaseHistidine-TaggedMaxiKit–HighYield 154
PrepEaseHistidine-TaggedMidiKit–HighSpecificity 153
PrepEaseHistidine-TaggedMidiKit–HighYield 154
PrepEaseHistidine-TaggedMiniKit–HighSpecificity 153
PrepEaseHistidine-TaggedMiniKit–HighYield 154
Histidine-TaggedProteinPurificationResins
PrepEaseHistidine-TaggedHighSpecificityPurificationResin 155
PrepEaseHistidine-TaggedHighYieldPurificationResin 155
L-Histidine,FreeBase 222
L-Histidine,Monohydrochloride,Monohydrate 222
hMLH1GenePromoterReporterVector 111
HNF-3bELISAKit 110
HNF-3bELISAKitwithNuclearExtractionKit 110
HNF4gCMVExpressionVector 114
HotStartPCR
HotStart-ITBindingProtein 21
HotStart-ITTaqDNAPolymerase 19
HotStart-ITTaqPCRMasterMix(2X) 20
HotStart-ITBindingProtein 21
HotStart-ITFideliTaqDNAPolymerase 17,25
HotStart-ITFideliTaqPCRMasterMix(2X) 18
HotStart-ITProbeOne-StepqRT-PCRMasterMixKit 44
HotStart-ITProbeqPCRMasterMix(2X) 38
HotStart-ITProbeqPCRMasterMixwithUDG(2X) 38
HotStart-ITSYBRGreenOne-StepqRT-PCRMasterMixKit 44
HotStart-ITSYBRGreenqPCRMasterMix(2X) 32
HotStart-ITSYBRGreenqPCRMasterMixwithUDG(2X) 32
HotStart-ITTaqDNAPolymerase 19
HotStart-ITTaqPCRMasterMix(2X) 20
HOXA2 25
HOXA2GenePromoterReporterVector 111
HOXA5GenePromoterReporterVector 111
H-ras 24
HSELuciferaseReporterVector 112
HTExoSAP-ITHigh-ThroughputPCRProductCleanup 50,149
HumanAngiogenesisAntibodyArray 133
HumanApurinic/ApyrimidinicEndonuclease1(APE1) 75
HumanCytokineAntibodyArray1.0 134
HumanCytokineAntibodyArray3.0 134
HumanGAPDHsiRNAControl 98
L-HydroxysuccinicAcid—SeeL-MalicAcid 227
I
IFNgGenePromoterReporterVector 111
IGRPGenePromoterReporterVector 111
IL1aGenePromoterReporterVector 111
IL2GenePromoterReporterVector 111
IL4 25
IL4GenePromoterReporterVector 111
Imidazole,Ultrapure 223
INFg 24
IPTG—SeeIsopropyl-b-D-Thiogalactopyranoside 223
IRF1CMVExpressionVector 114
IRF1LuciferaseReporterVector 112
IRF7 25
IRF7GenePromoterReporterVector 111
IsocitrateDehydrogenase,(NADP+Specific),Recombinant 223
DL-Isoleucine 223
L-Isoleucine 223
Isopropyl-b-D-Thiogalactopyranoside(IPTG),DioxaneFree,Ultrapure 223
ISRELuciferaseReporterVector 112
J
JUNCMVExpressionVector 114
JUNBGenePromoterReporterVector 111
K
KanamycinSulfate 223
a-KetoglutaricAcid,DisodiumSalt,Dihydrate 224
a-KetoglutaricAcid,FreeAcid 224
Kinases
OptiKinase 68
T4PolynucleotideKinase(PNK) 69
KlenowDNAPolymeraseI 87
L
Labeling
SequenaseRandomPrimerLabelingKit 127
L-(+)-LacticAcid,LithiumSalt 224
Lactoferrin 224
Product Name Index | Alphabetical
279For bulk or alternate pack sizes, email us at [email protected].
Product Page Product Page
D-(+)-Lactose,Monohydrate 224
N-LauroylSarcosine,SodiumSalt,Ultrapure 224
LaurylSulfate,LithiumSalt—SeeLithiumDodecylSulfate 225
LaurylSulfate,SodiumSalt—SeeSodiumDodecylSulfate 240
LBAgar,Ready-MadePowder 224
LBBroth,Ready-MadePowder 224
Lecithin 224
LeptomycinB 224
DL-Leucine 225
L-Leucine 225
Leupeptin,Ultrapure 225
Ligases
E. coliDNALigase 70
T4DNALigase 70
T4RNALigase 71
T4RNALigase2 71
Ligate-ITRapidLigationKit 125
DL-a-LipoicAcid 225
LithiumChloride(LiCl),7.5MSolution,Ultrapure 225
LithiumDodecylSulfate,Ultrapure 225
LithiumLactate—SeeL-(+)-LacticAcid,LithiumSalt 224
LithiumLaurylSulfate—SeeLithiumDodecylSulfate 225
LongandAccuratePCR
FideliTaqDNAPolymerase 15
FideliTaqPCRMasterMix(2X) 16
HotStart-ITFideliTaqDNAPolymerase 17
HotStart-ITFideliTaqPCRMasterMix(2X) 18
LowMolecularWeightMarker,10-100nt 132,165
LuciferaseReporterVectors
TA-luc(EmptyControl)LuciferaseReporterVector 112
AP1(1)LuciferaseReporterVector 112
AP1(2)LuciferaseReporterVector 112
AP3LuciferaseReporterVector 112
ARLuciferaseReporterVector 112
CRE(1)LuciferaseReporterVector 112
E2F(1)LuciferaseReporterVector 112
E2F(2)LuciferaseReporterVector 112
E2F1(3)LuciferaseReporterVector 112
ELK1LuciferaseReporterVector 112
ERLuciferaseReporterVector 112
GAS/ISRELuciferaseReporterVector 112
GASLuciferaseReporterVector 112
GATALuciferaseReporterVector 112
GATA1/GATA2LuciferaseReporterVector 112
GATA1LuciferaseReporterVector 112
GATA2LuciferaseReporterVector 112
GATA3LuciferaseReporterVector 112
GATA4LuciferaseReporterVector 112
GRLuciferaseReporterVector 112
HIF1LuciferaseReporterVector 112
HSELuciferaseReporterVector 112
IRF1LuciferaseReporterVector 112
ISRELuciferaseReporterVector 112
MEF1LuciferaseReporterVector 112
MEF2LuciferaseReporterVector 112
MEF3LuciferaseReporterVector 112
NF-ATLuciferaseReporterVector 112
NFkB(1)LuciferaseReporterVector 112
NFkB(2)LuciferaseReporterVector 112
p53LuciferaseReporterVector 112
PRLuciferaseReporterVector 112
RARLuciferaseReporterVector 112
RXRLuciferaseReporterVector 112
RXR(2)LuciferaseReporterVector 112
SmadLuciferaseReporterVector 112
Smad3/Smad4LuciferaseReporterVector 112
Sp1LuciferaseReporterVector 112
SP1(2)LuciferaseReporterVector 112
SRELuciferaseReporterVector 112
SRFLuciferaseReporterVector 112
STAT1p84/p91LuciferaseReporterVector 112
STAT1(2)LuciferaseReporterVector 112
STAT3(1)LuciferaseReporterVector 112
STAT4LuciferaseReporterVector 112
VDRLuciferaseReporterVector 112
YY1LuciferaseReporterVector 112
LuriaAgar,Ready-MadePowder 225
LuriaBroth,Ready-MadePowder 225
L-Lysine,Hydrochloride 226
Lysozyme,Ultrapure 226
Product Name Index | Alphabetical
280 888-362-2447 | 216-765-5000 | usb.affymetrix.com
Product Page Product Page
M
M13mp18,Single-Stranded,0.2µg/ul 129
M13mp18DNA,Single-Stranded,1.0µg/µl 129
MAFKCMVExpressionVector 114
MagnesiumChloride(MgCl2),1MSolution,Ultrapure 226
MagnesiumChloride,Hexahydrate,Ultrapure 226
MagniTaqMultiplexPCRMasterMix(2X) 22
MalateDehydrogenase,Lyophilized 226
MaleicAcid 226
DL-MalicAcid 226
L-MalicAcid 227
Maltose,Monohydrate 227
D-(+)-Maltose,Monohydrate 227
MammalianFull-lengthExpressionVectors
ATF1CMVExpressionVector 114
ATF3CMVExpressionVector 114
BTG2CMVExpressionVector 114
CDX2CMVExpressionVector 114
CITED1CMVExpressionVector 114
E2F2CMVExpressionVector 114
ERaCMVExpressionVector 114
FOSCMVExpressionVector 114
GATA1CMVExpressionVector 114
GFI1CMVExpressionVector 114
GR(NR3C1)CMVExpressionVector 114
HNF4gCMVExpressionVector 114
IRF1CMVExpressionVector 114
JUNCMVExpressionVector 114
MAFKCMVExpressionVector 114
MAXCMVExpressionVector 114
MYCCMVExpressionVector 114
NF-ATC1CMVExpressionVector 114
NFkBp50CMVExpressionVector 114
NFkBp65(RelA)CMVExpressionVector 114
NF-YCCMVExpressionVector 114
NR0B1CMVExpressionVector 114
NR1I2CMVExpressionVector 114
NR1I3CMVExpressionVector 114
p53CMVExpressionVector 114
PBX1CMVExpressionVector 114
PPARaCMVExpressionVector 114
PPARb(PPARΔ)CMVExpressionVector 114
PPARg-1CMVExpressionVector 114
PPARg-2CMVExpressionVector 114
PREBCMVExpressionVector 114
SMAD1(MADH1)CMVExpressionVector 114
SRYCMVExpressionVector 114
STAT1CMVExpressionVector 114
STAT5BCMVExpressionVector 114
USF1CMVExpressionVector 114
Mannitol 227
D-(+)-Mannose 227
Markers
DNAMarkers 130
OligonucleotideMarker 132
ProteinMarkers 136
MAXCMVExpressionVector 114
MDR1GenePromoterReporterVector 111
MEF1LuciferaseReporterVector 112
MEF-2ColdControlProbe 110
MEF-2ControlNuclearExtract 110
MEF-2ELISAKit 110
MEF-2ELISAKitwithNuclearExtractionKit 110
MEF2LuciferaseReporterVector 112
MEF3LuciferaseReporterVector 112
MES,Anhydrous 227
MES,Monohydrate,Ultrapure 227
MES,SodiumSalt 227
5-Methyl-2’-Deoxycytidine-5’-Monophosphate,DisodiumSalt 228
MethyleneBlue,Trihydrate 228
N,N’-Methylene-bis-Acrylamide,Ultrapure 228
4-Methylumbelliferyl-a-L-Iduronide,SodiumSalt 228
MicrococcalNuclease 81
MineralOil,Ultrapure 228
M-MLVReverseTranscriptase 90,162
MonosodiumGlutamate—SeeGlutamicAcid,MonosodiumSalt 218
MOPS,Ultrapure 228
MouseAngiogenesisAntibodyArray 133
MouseCytokineAntibodyArray1.0 134
Product Name Index | Alphabetical
281For bulk or alternate pack sizes, email us at [email protected].
Product Page Product Page
mRNAAssay
Poly(A)Tail-LengthAssayKit 158
MTT,Ultrapure 228
MultiplexPCR
HotStart-ITFideliTaqDNAPolymerase 25
MagniTaqMultiplexPCRMasterMix(2X) 22
PromoterMethylationPCRKit 24
Mutagenesis
Change-ITMultipleMutationSiteDirectedMutagenesisKit 126
MYCCMVExpressionVector 114
MyoDGenePromoterReporterVector 111
N
b-NAD—SeeNicotinamideAdenineDinucleotide 229
NADH—Seeb-NicotinamideAdenineDinucleotide,Reduced 230
NADP—SeeNicotinamideAdenineDinucleotidePhosphate 230
NADPH—SeeNicotinamideAdenineDinucleotidePhosphate,Reduced 230
a-NaphthylPhosphate,MonosodiumSalt 229
NBT—Seep-NitroBlueTetrazoliumChloride 230
Neocuproine,Hydrochloride 229
NeomycinSulfate,Ultrapure 229
NES 25
NeutralRed 229
NextGenerationSequencing(NGS)LibraryPrep
Prep2SeqDNALibraryPrepKitforIllumina 123,170
Prep2SeqMultiplexOligoAdaptersforIlluminaplatform 124,171
NF-ATLuciferaseReporterVector 112
NF-ATC1CMVExpressionVector 114
NFATHeLaReporterStableCellLine 122
NFATK562ReporterStableCellLine 122
NFkB293ReporterStableCellLine 122
NFkBA549ReporterStableCellLine 122
NFkBC2C12ReporterStableCellLine 122
NFkBc-RelELISAKit 110
NFkBc-RelELISAKitwithNuclearExtractionKit 110
NFkBControlNuclearExtract 110
NFkBColdControlProbe 110
NFkBHeLaReporterStableCellLine 122
NFkB(1)LuciferaseReporterVector 112
NFkB(2)LuciferaseReporterVector 112
NFkBNIH3T3ReporterStableCellLine 122
NFkBp50CMVExpressionVector 114
NFkBp50ELISAKit 110
NFkBp50ELISAKitwithNuclearExtractionKit 110
NFkBp50KnockdownHeLaStableCellLine 122
NFkBp50-TFInteractionArrayI 118
NFkBp50-TFInteractionArrayII 118
NFkBp50-TFInteractionArrayIII 118
NFkBp52ELISAKit 110
NFkBp52ELISAKitwithNuclearExtractionKit 110
NFkBp65ELISAKit 110
NFkBp65ELISAKitwithNuclearExtractionKit 110
NFkBp65(RelA)CMVExpressionVector 114
NFkBp65-TFInteractionArrayI 118
NFkBp65-TFInteractionArrayII 118
NFkBp65-TFInteractionArrayIII 118
NFkBRel-bELISAKit 110
NFkBRel-bELISAKitwithNuclearExtractionKit 110
NF-YCCMVExpressionVector 114
Niacinamide 229
Nicotinamide—SeeNiacinamide 229
b-NicotinamideAdenineDinucleotide 229
b-NicotinamideAdenineDinucleotide,LithiumSalt,Dihydrate 229
b-NicotinamideAdenineDinucleotide,MonosodiumSalt 230
b-NicotinamideAdenineDinucleotidePhosphate,Monosodium,Tetrahydrate 230
b-NicotinamideAdenineDinucleotidePhosphate-Reduced,Tetrasodium,Tetrahydrate230
b-NicotinamideAdenineDinucleotide-Reduced,Disodium,Trihydrate 230
NIS 25
p-NitroBlueTetrazoliumChloride,Ultrapure 230
4-Nitrophenyl-N-Acetyl-b-D-Galactosaminide 230
4-Nitrophenyl-a-L-Fucopyranoside 231
4-Nitrophenyl-a-D-Galactopyranoside 231
4-Nitrophenyl-b-D-Glucopyranoside 231
4-Nitrophenyl-b-D-Glucuronide 231
4-Nitrophenyl-a-D-Maltoside 231
4-Nitrophenyl-b-D-Mannopyranoside 231
4-Nitrophenyl-b-D-Xylopyranoside 231
NME2 25
Non-SpecificNucleases
MicrococcalNuclease 81
PhosphodiesteraseI 81
Product Name Index | Alphabetical
282 888-362-2447 | 216-765-5000 | usb.affymetrix.com
Product Page Product Page
NR0B1CMVExpressionVector 114
NR1I2CMVExpressionVector 114
NR1I3CMVExpressionVector 114
NuclearExtractionKit 102
NuclearReceptorProtein/DNAArray 115
NuclearReceptorProtein/DNAArrayRefill 115
Nucleases
DNaseI,Lyophilized 72
DNaseI,Solution 72
rDNaseI,RNase-Free 73
10XDNaseIBuffer 74
ExonucleaseI 76
ExonucleaseIII 77
ExonucleaseVII 77
HumanApurinic/ApyrimidinicEndonuclease1(APE1) 75
MicrococcalNuclease 81
PhosphodiesteraseI 81
RNaseI 79
RnaseI'A',Powder 79
RNaseA,Lyophilized 79
RNaseA,MBGrade,Solution 80
RNaseH 80
ShrimpDNase,Recombinant 74
T4EndonucleaseVII 75
T7Gene6Exonuclease 78
Nucleoside-5’-Triphosphates(ATP,CTP,GTP,UTP),SetofFour,100mM 61,231Solutions,Ultrapure,4x25µmol(250µl)eachNTPperpack
NucleotideLabelingReagents
Biotin-11-dXTPAnalog(DNALabelingReagent,DLR) 57
Biotin-11-ψTPAnalog(RNALabelingReagent,RLR) 57
NucleotideSolutions
Adenosine-5’-Triphosphate(ATP) 189
2’-Deoxyadenosine-5’-Triphosphate(dATP) 210
2’-Deoxycytidine-5’-Triphosphate(dCTP) 210
2’-Deoxyguanosine-5’-Triphosphate(dGTP) 210
2’-Deoxynucleoside-5’-Triphosphates(dNTP),SetofFour 211
2’-Deoxythymidine-5’-Triphosphate(dTTP) 211
2’-Deoxyuridine-5’-Triphosphate(dUTP) 211
2’,3’-Dideoxyadenosine-5’-Triphosphate(ddATP) 212
2’,3’-Dideoxycytidine-5’-Triphosphate(ddCTP) 212
2’,3’-Dideoxythymidine-5’-Triphosphate(ddTTP) 212
Guanosine-5’-Triphosphate(GTP) 221
PCR-NucleotideMixes 232
NZCYMBroth,Ready-MadePowder 232
O
Oligo(dT)12-18Primer,MW≅4500 129,231
Oligop(dT)Cellulose,Ultrapure 125
Oligop(dT)12-18SodiumSalt 129,232
OligonucleotideMarker
LowMolecularWeightMarker,10–100nt 132
One-StepRT-PCRKit 47
OptiKinase 68
Ovalbumin—SeeAlbumin,Egg 199
P
p53CMVExpressionVector 114
p53ColdControlProbe 110
p53ELISAKit 110
p53ELISAKitwithNuclearExtractionKit 112
p53LuciferaseReporterVector 112
PABA—Seep-AminoBenzoicAcid 199
Pancreatin1X 232
ParaformaldehydeSolution,4%inPBS 232
PassiveReferenceDyes
FluoresceinPassiveReferenceDye 53
ROXPassiveReferenceDye 53
PBS,10XSolution,pH7.4,Ultrapure 232
PBX1CMVExpressionVector 114
PCRAmplification
EndPointPCR 11
HotStartPCR 19
LongandAccuratePCR 15
MultiplexPCR 22
Real-TimeqPCR 26
Real-TimeReverseTranscriptionPCR(qRT-PCR) 39
ReverseTranscriptionRT-PCR—One-StepMasterMix 45
ReverseTranscriptionRT-PCR—Kits 47
PCRCleanup
ExoSAP-ITPCRProductCleanup 49,148
HTExoSAP-ITHigh-ThroughputPCRProductCleanup 50,149
Product Name Index | Alphabetical
283For bulk or alternate pack sizes, email us at [email protected].
Product Page Product Page
PCRMarkers,50-2000bp 130
PCRNucleotideMix,10mMSolution,Ultrapure 60,232
PCRNucleotideMix,25mMSolution,Ultrapure 60,232
PCRNucleotideMixwithdUTP,10mMSolution,Ultrapure 60,232
PCRPrimers(forPromoterMethylationPCRKit) 24
14-3-3 24
ABL1 24
BAGE 24
BRCA1 24
CalcitoninCGPR 24
CASP8 25
CD14 24
CDKN2A 24
COX2 24
CyclinD2 24
DAPK 24
GAGE1 24
glut4 24
H-ras 24
HOXA2 25
IL4 25
INFg 24
IRF7 25
NES 25
NIS 25
NME2 25
PDGFB 25
POMC 25
PRA2 25
Survivin 25
TFF1 25
VHL 25
PCRProductPre-SequencingKit 52
PCRPurification
ExoSAP-ITPCRProductCleanup 49,148
HTExoSAP-ITHigh-ThroughputPCRProductCleanup 50,149
PCRProductPre-SequencingKit 52
PrepEaseDNACleanupKits 52,146
PrepEaseGelExtractionKits 52,146
PrepEasePCRPurification96-WellPlateKits(Ultrafiltration) 53,148
SBECleanupReagent 52
PDGFB 25
PEG
PolyethyleneGlycol400 235
PolyethyleneGlycol6000 235
PolyethyleneGlycol8000 235
PenicillinG,Potassium 232
Pepsin 232
Pepsin1:10,000 233
PepstatinA,Ultrapure 233
Peptones
BacteriologicalPeptone 202
CaseinHydrolysate 205
YeastExtractandHydrolysate 251
Phenol,pH8.0,Equilibrated,Ultrapure 233
Phenol,Ultrapure 233
Phenol:Chloroform:IsoamylAlcohol(25:24:1),Ultrapure 233
PhenolRed,SodiumSalt 234
Phenolphthalein 234
Phenolsulfonphthalein—SeePhenolRed 234
DL-Phenylalanine 234
L-Phenylalanine 234
PhenylmethylSulfonylFluoride,Ultrapure 234
Phosphatases
CalfIntestinalAlkalinePhosphatase(CIAP) 82
ShrimpAlkalinePhosphatase(SAP) 83
SuperSAPShrimpAlkalinePhosphatase 84
Phosphocreatine—SeeCreatinePhosphate 208
PhosphodiesteraseI 81
PIPES 208
PIPES,DisodiumSalt 234
PlasmidPurification-BAC
PrepEaseBACPurificationKit 145
PlasmidPurification-Endotoxin-FreePreps
PrepEaseEndotoxin-FreeMaxiPlasmidKit 142
PlasmidPurification-HighThroughput
PrepEasePlasmid96-wellPlateKit 144
PrepeasePlasmid96-wellPlateCoreKit 144
Product Name Index | Alphabetical
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Product Page Product Page
PlasmidPurification-MediumThroughput
PrepEasePlasmid8-wellStripKit 143
PrepEase8-wellStripStarterKitforVacuum 143
PrepEaseVacuumManifold 143
PrepEasePurification-MiditoGigaPreps
PrepEaseMaxiPlasmidKit 140
PrepEaseMidiPlasmidKit 140
PrepEaseGigaPlasmidKit 140
PlasmidPurification-SpinColumns
PrepEaseMiniSpinPlasmidKit 141
PrepEaseQuickMiniSpinPlasmidKit 141
PlasmidPurification-Yeast
PrepEaseYeastPlasmidIsolationKit 145
PMSF—SeePhenylmethylSulfonylFluoride 234
PNK
OptiKinase 68
T4PolynucleotideKinase 69
Poly(A)Polymerase,Yeast 91,159
Poly(A)Tail-LengthAssayKit 158
PolyanetholesulfonicAcid,SodiumSalt 234
Poly(dI-dC)•Poly(dI-dC)SodiumSalt 235
PolyethyleneGlycol400 235
PolyethyleneGlycol6000,Flakes 235
PolyethyleneGlycol8000,Flakes,Ultrapure 235
PolyethyleneGlycol8000,Powder 235
Polymerases
AMVReverseTranscriptase 90,162
DNAPolymeraseI 87
E. coliRNAPolymeraseHoloenzyme 91,159
Exonuclease-FreeKlenow 86
Exonuclease-FreeKlenowTrisBuffer 86
KlenowDNAPolymeraseI 87
M-MLVReverseTranscriptase 90,162
Poly(A)Polymerase,Yeast 91,159
SequenaseVersion2.0DNAPolymerase 88,174
T3RNAPolymerase 92,160
T7DNAPolymerase 88
T7RNAPolymerase 92,161
TthDNAPolymerase 89
TerminalDeoxynucleotidylTransferase,Recombinant(rTdT) 93
PolymixinBSulfate 235
PolyvinylPyrrolidone(PVP)—SeePVP-K-30 237
POMC 25
POMCGenePromoterReporterVector 111
PonceauS,SodiumSalt,Ultrapure 236
PotassiumChloride,Ultrapure 236
PotassiumChloride(KCl),2MSolution,Ultrapure 236
PotassiumPhosphate,Dibasic 236
PotassiumPhosphate,Dibasic,Ultrapure 236
PotassiumPhosphate,Monobasic 236
PotassiumPhosphate,Monobasic,Anhydrous,Ultrapure 236
PPARColdControlProbe 110
PPARaCMVExpressionVector 114
PPARaColdControlProbe 110
PPARaControlCos1NuclearExtract 110
PPARaELISAKit 110
PPARaELISAKitwithNuclearExtractionKit 110
PPARa-TFInteractionArrayI 118
PPARa-TFInteractionArrayII 118
PPARa-TFInteractionArrayIII 118
PPARb(PPARΔ)CMVExpressionVector 114
PPARg-1CMVExpressionVector 114
PPARg-2CMVExpressionVector 114
PPARg-TFInteractionArrayI 118
PPARg-TFInteractionArrayII 118
PPARg-TFInteractionArrayIII 118
PRLuciferaseReporterVector 112
PRA2 25
PREBCMVExpressionVector 114
Prep2SeqDNALibraryPrepKitforIlluminaplatform 123,170
Prep2SeqMultiplexOligoAdaptersforIlluminaplatform 124,171(Containsadaptersequences1-12)
Prep2SeqMultiplexOligoAdaptersforIlluminaplatform 124,171(Containsadaptersequences13-27)
PrepEase8-wellStripStarterKitforVacuum 143
PrepEaseBACPurificationKit 145
PrepEaseDNACleanupKits 146
PrepEaseEndotoxin-FreeMaxiPlasmidKit 142
PrepEaseGelExtractionKits 146
PrepEaseGenomicDNAIsolationKits 147
Product Name Index | Alphabetical
285For bulk or alternate pack sizes, email us at [email protected].
Product Page Product Page
PrepEaseGigaPlasmidKit 140
PrepEaseHistidine-TaggedHighSpecificityPurificationResin 155
PrepEaseHistidine-TaggedHighYieldPurificationResin 155
PrepEaseHistidine-TaggedProteinPurificationMaxiKit-HighSpecificity 153
PrepEaseHistidine-TaggedProteinPurificationMaxiKit-HighYield 154
PrepEaseHistidine-TaggedProteinPurificationMidiKit-HighSpecificity 153
PrepEaseHistidine-TaggedProteinPurificationMidiKit-HighYield 154
PrepEaseHistidine-TaggedProteinPurificationMiniKit-HighSpecificity 153
PrepEaseHistidine-TaggedProteinPurificationMiniKit-HighYield 154
PrepEaseMaxiPlasmidKit 140
PrepEaseMidiPlasmidKit 140
PrepEaseMiniSpinPlasmidKit 141
PrepEasemRNAMiniSpinKit 150
PrepEasePCRPurification96-WellPlateKits(Ultrafiltration) 148
PrepEasePlasmid8-wellStripKit 143
PrepEasePlasmid96-wellPlateCoreKit 144
PrepEasePlasmid96-wellPlateKit 144
PrepEaseProteinPurificationGlutathioneAgarose4B 155
PrepEaseQuickMiniSpinPlasmidKit 141
PrepEaseRNASpinKits 150
PrepEaseRNA/ProteinSpinKit 151
PrepEaseSequencingDyeCleanupKits 152
PrepEaseVacuumManifold 143
PrepEaseYeastPlasmidIsolationKit 145
PromoterMethylationPCRKit 24
ProteinAnalysis
AngiogenesisAntibodyArrays 133
CancerAntigenELISAKits 134
CytokineAntibodyArrays 134
ProteinMarkers
IEFProteinMarkers 134
ProteinMarkers,10-225kDa 134
ProteinMarkers,10-225kDa 136
Protein/DNAArrayI 116
Protein/DNAArrayIRefillKit 116
Protein/DNAArrayII 116
Protein/DNAArrayIIRefillKit 116
Protein/DNAArrayIII 116
Protein/DNAArrayIIIRefillKit 116
Protein/DNAArrayIV 116
Protein/DNAArrayIVRefillKit 116
Protein/DNAArrayV 116
Protein/DNAArrayVRefillKit 116
Proteinases
ProteinaseK 137
ProteinaseKSolution 137
ProteinaseK 137,237
ProteinaseKSolution 137
pUC19DNA—SeeMolecularBiologyProducts&Kitssection 129
pUC19DNA,0.2µg/µl 129
pUC19DNA,1.0µg/µl 129
PurificationSolutions
Phenol,pH8.0,Equilibrated 233
Phenol:Chloroform:IsoamylAlcohol(25:24:1) 233
PVP-K-30 237
Pyrophosphatase
Pyrophosphatase,Inorganic(Recombinant)(rPPase) 85
PyruvicAcid,SodiumSalt 237
R
D-(+)-Raffinose,Pentahydrate 237
RapidGel40%LiquidAcrylamideStockSolution,Ultrapure 237
RapidGel-XL6%LiquidAcrylamideDNASequencingGel,TBEFormulation,Ultrapure 238
RapidRunAgaroseBuffer,20XSolution,Ultrapure 238
RapidRunLoadingDye,Ultrapure 238
RARLuciferaseReporterVector 112
RB1GenePromoterReporterVector 111
Real-TimeqPCR
HotStart-ITProbeqPCRMasterMix(2X) 38
HotStart-ITProbeqPCRMasterMixwithUDG(2X) 38
HotStart-ITSYBRGreenqPCRMasterMix(2X) 32
HotStart-ITSYBRGreenqPCRMasterMixwithUDG(2X) 32
VeriQuestFastProbeqPCRMasterMix(2X) 36
VeriQuestFastProbeqPCRMasterMix,NoReferenceDye(2X) 37
VeriQuestFastSYBRGreenqPCRMasterMix(2X) 30
VeriQuestFastSYBRGreenqPCRMasterMixwithFluorescein 31
VeriQuestProbeqPCRMasterMix(2X) 33
VeriQuestProbeqPCRMasterMix,NoReferenceDye(2X) 35
VeriQuestSYBRGreenqPCRMasterMix(2X) 27
VeriQuestSYBRGreenqPCRMasterMixwithFluorescein(2X) 29
VeriQuestTaqDNAPolymerase 26
Product Name Index | Alphabetical
286 888-362-2447 | 216-765-5000 | usb.affymetrix.com
Product Page Product Page
Real-TimeReverseTranscriptionPCR(qRT-PCR)
First-StrandcDNASynthesisKitforReal-TimePCR 39
HotStart-ITProbeOne-StepqRT-PCRMasterMixKit 44
HotStart-ITSYBRGreenOne-StepqRT-PCRMasterMixKit 44
VeriQuestProbeOne-StepqRT-PCRMasterMix 42
VeriQuestProbeOne-StepqRT-PCRMasterMix,NoReferenceDye(2X) 43
VeriQuestSYBRGreenOne-StepqRT-PCRMasterMix(2X) 40
VeriQuestSYBRGreenOne-StepqRT-PCRMasterMixwithFluorescein(2X) 41
RecAProtein 65
ReporterStableCellLines
AP-1293ReporterStableCellLine 122
CHO/GFP-GR*StableCellLine 122
CHO/GFP-NFkBp65StableCellLine 122
CREB293ReporterStableCellLine 122
CREBCHOReporterStableCellLine 122
GR293ReporterStableCellLine 122
GRHeLaReporterStableCellLine 122
HIFNIH3T3ReporterStableCellLine 122
NFATHeLaReporterStableCellLine 122
NFATK562ReporterStableCellLine 122
NFkB293ReporterStableCellLine 122
NFkBA549ReporterStableCellLine 122
NFkBC2C12ReporterStableCellLine 122
NFkBHeLaReporterStableCellLine 122
NFkBNIH3T3ReporterStableCellLine 122
NFkBp50KnockdownHeLaStableCellLine 122
SRFHeLaReporterStableCellLine 122
STAT-1HeLaReporterStableCellLine 122
TAD-KinaseReporterStableCellLine 122
ReverseTranscriptases
AMVReverseTranscriptase 90,162
M-MLVReverseTranscriptase 90,162
ReverseTranscriptionRT-PCR-Kits
First-StrandcDNASynthesisKitforReal-TimePCR 47
One-StepRT-PCRKit 47
Two-StepRT-PCRKit 48
ReverseTranscriptionRT-PCR-One-StepMasterMix
FideliTaqRT-PCRMasterMix(2X) 46
RT-PCRMasterMix(2X) 45
Riboflavin 238
D-(–)-Ribose 238
Rifampin 238
RNA,FreeAcid 238
RNA-ReverseTranscription
AMVReverseTranscriptase 90,162
M-MLVReverseTranscriptase 90,162
RNAPolymerases
E. coliRNAPolymeraseHoloenzyme 91,159
Poly(A)Polymerase,Yeast 91,159
T3RNAPolymerase 92,160
T7RNAPolymerase 92,161
RNaseInhibitors
RNaseInhibitor(HumanPlacenta) 94,164
RNaseInhibitor(Recombinant) 94,164
RNAPurification
PrepEasemRNAMiniSpinKit 150
PrepEaseRNA/ProteinSpinKit 151
PrepEaseRNASpinKits 150
RNATranscription
E. coliRNAPolymeraseHoloenzyme 91,159
Poly(A)Polymerase,Yeast 91,159
T3RNAPolymerase 92,160
T7RNAPolymerase 92,161
RNaseI 79,163
RNaseI‘A’,Powder 79,163
RNaseA,Lyophilized 79,163,239
RNaseA,MolecularBiologyGrade,Solution 80,163
RNaseH 80,163
RNaseInhibitor(HumanPlacenta) 94,164
RNaseInhibitor(Recombinant) 94,164
RNases
RNaseI 79
RnaseI'A'Powder 79
RNaseA,Lyophilized 79
RNaseA,MBGrade,Solution 80
RNaseH 80
ROXPassiveReferenceDye 53
RT-PCRMasterMix(2X) 45
Product Name Index | Alphabetical
287For bulk or alternate pack sizes, email us at [email protected].
Product Page Product Page
RubyTaqDNAPolymerase 13
RubyTaqPCRMasterMix(2X) 14
RXRLuciferaseReporterVector 112
RXR(2)LuciferaseReporterVector 112
S
SAMe-PTS—SeeS-Adenosyl-L-MethionineSulfatep-Toluenesulfonate 190
SBECleanupReagent 52
SDS—SeeSodiumDodecylSulfate 240
SequenaseEnzyme&Kits
SequenasePCRProductSequencingKit 176
SequenaseQuickDenaturePlasmidDNASequencingKit 177
SequenaseVersion2.0DNAPolymerase 88,174
SequenaseVersion2.0DNASequencingKit 175
SequenasePCRProductSequencingKit 176
SequenaseQuickDenaturePlasmidDNASequencingKit 177
SequenaseRandomPrimerLabelingKit 127
SequenaseVersion2.0DNAPolymerase 88,174
SequenaseVersion2.0DNAPolymerasewithPyrophosphatase 85
SequenaseVersion2.0DNASequencingKit 175
SequencingDyeCleanup
PrepEaseSequencingDyeCleanupKits 152
DL-Serine(Non-Animal) 239
ShrimpAlkalinePhosphatase(SAP) 83
ShrimpDNase,Recombinant 74
Single-StrandedDNABindingProtein(SSB) 65
SmadLuciferaseReporterVector 112
SMAD1(MADH1)CMVExpressionVector 114
SMAD-2ELISAKit 110
SMAD-2ELISAKitwithNuclearExtractionKit 110
Smad3/Smad4LuciferaseReporterVector 112
SMAD-4ELISAKit 110
SMAD-4ELISAKitwithNuclearExtractionKit 110
SodiumAcetate(NaOAc),3MSolution,pH5.5,Ultrapure 239
SodiumAspartate—SeeAsparticAcid,SodiumSalt 201
SodiumAzide,Ultrapure 239
SodiumCacodylate—SeeCacodylicAcid,SodiumSalt 204
SodiumCarbonate,Anhydrous 239
SodiumCaseinate—SeeCasein,Sodium 205
SodiumChloride,Ultrapure 239
SodiumChloride(NaCl),5MSolution,Ultrapure 239
SodiumCitrate—SeeCitricAcid,TrisodiumSalt 207
SodiumDeoxycholate—SeeDeoxycholicAcid,SodiumSalt 210
SodiumDodecylSulfate(SDS) 240
SodiumDodecylSulfate(SDS),10%Solution,Ultrapure 240
SodiumDodecylSulfate(SDS),20%Solution,Ultrapure 240
SodiumDodecylSulfate(SDS),Ultrapure 240
SodiumGlutamate—SeeGlutamicAcid,MonosodiumSalt 218
SodiumLaurylSulfate—SeeSodiumDodecylSulfate 240
Sodium-N-LaurylSarcosine—SeeN-LaurylSarcosine,SodiumSalt 224
SodiumPhosphate,DibasicAnhydrous 240
SodiumPhosphate,Dibasic,Heptahydrate,Ultrapure 240
SodiumPhosphate,Monobasic 240
SodiumPhosphate,Monobasic,Monohydrate,Ultrapure 241
SodiumPyruvate—SeePyruvicAcid,SodiumSalt 237
SodiumSelenite 241
SolubleStarch—SeeStarch-Soluble 242
SodiumSuccinate—SeeSuccinicAcid,DisodiumSalt 242
D-Sorbitol,Ultrapure 241
SP-1ColdProbe 110
SP-1ControlRecombinantProtein 110
SP-1ELISAKit 110
SP-1ELISAKitwithNuclearExtractionKit 110
Sp1LuciferaseReporterVector 112
SP1(2)LuciferaseReporterVector 112
Spermidine,Trihydrochloride,Ultrapure 241
Spermine,Tetrahydrochloride 241
SRFHeLaReporterStableCellLine 122
SRELuciferaseReporterVector 112
SRFLuciferaseReporterVector 112
SRYCMVExpressionVector 114
SSC,20XSolution,Ultrapure 241
SSPE,20XSolution,Ultrapure 241
Starch,Hydrolyzed,Ultrapure 242
Starch-Soluble,Ultrapure 242
STAT-1ColdControlProbe 110
STAT-1ControlMCF7(IFNg) 110
STAT-1ELISAKit 110
STAT-1ELISAKitwithNuclearExtractionKit 110
Product Name Index | Alphabetical
288 888-362-2447 | 216-765-5000 | usb.affymetrix.com
Product Page Product Page
STAT-1HeLaReporterStableCellLine 122
STAT1CMVExpressionVector 114
STAT1p84/p91LuciferaseReporterVector 112
STAT1GenePromoterReporterVector 111
STAT1(2)LuciferaseReporterVector 112
STAT3(1)LuciferaseReporterVector 112
STAT4LuciferaseReporterVector 112
STAT5BCMVExpressionVector 114
StockSolutions
AmmoniumAcetate(NH4OAc),5MSolution 199
Betaine,5MSolution 202
Dithiothreitol(DTT)0.1MSolution 213
EDTA,0.5MSolution 215
LithiumChloride(LiCl),7.5MSolution 225
MagnesiumChloride(MgCl2),1MSolution 226
PotassiumChloride(KCl),2MSolution 236
SodiumAcetate(NaOAc),3MSolution,pH5.5 239
SodiumChloride(NaCl),5MSolution 239
SodiumDodecylSulfate(SDS),10%Solution 240
SodiumDodecylSulfate(SDS),20%Solution 240
Water,Nuclease-Free 250
Water,RNase-Free,DEPCTreated 250
StopSolution,Ultrapure 242
Streptavidin,Ultrapure 242
Streptavidin-AlkalinePhosphataseConjugate 242
StreptomycinSulfate,Ultrapure 242
SuccinicAcid,DisodiumSalt,Hexahydrate 242
SuccinicAcid,FreeAcid 242
Sucrose 243
Sucrose,Crystalline 243
Sucrose,Ultrapure 243
SulfanilicAcid 243
SuperSAPShrimpAlkalinePhosphatase 84
Survivin 25
SURVIVINGenePromoterReporterVector 111
T
T3PromoterPrimer 178
T3RNAPolymerase,Highconcentration,200units/µl 92,160
T3RNAPolymerase,Standardconcentration,20units/µl 92,160
T4DNALigase,Highconcentration,10units/µl 70
T4DNALigase,Standardconcentration,1unit/µl 70
T4EndonucleaseVII 75
T4Gene32Protein,Highconcentration,≥10µg/µl 66
T4Gene32Protein,Standardconcentration,5µg/µl 66
T4PolynucleotideKinase(PNK) 69
T4RNALigase 71
T4RNALigase2 71
T7DNAPolymerase 88
T7Gene6Exonuclease 78
T7RNAPolymerase,Highconcentration,>80units/µl 92,161
T7RNAPolymerase,Standardconcentration,20units/µl 92,161
TA-luc(EmptyControl)LuciferaseReporterVector 112
TAD-KinaseReporterStableCellLine 122
TAEBuffer,10XSolution,Ultrapure 243
TAEBuffer,50XSolution,Ultrapure 243
TaqDNAPolymerase 11
TaqPCRKit 12
TaqPCRMasterMix(2X) 12
Taurine,Ultrapure 243
TBEBuffer,5XSolution,Ultrapure 243
TBEBuffer,10XReady-MixedPowder,Ultrapure 243
TBS,20XSolution,pH7.4,Ultrapure 244
TBSwithTween(TBST),20XSolution,pH7.4,Ultrapure 244
TCEP-HCI,0.5MSolution,pH6.6±0.1 244
TEBuffer,1XSolution,pH7.0 244
TEBuffer,1XSolution,pH7.0,LowEDTA 244
TEBuffer,1XSolution,pH8.0 244
TEBuffer,1XSolution,pH8.0,LowEDTA 244
TEBuffer,1XSolution,Ultrapure 244
TEBuffer,50XSolution,Ultrapure 244
TEMED—SeeTetramethylethylenediamine 245
TerrificBroth,Ready-MadePowder 245
TerminalDeoxynucleotidylTransferase,Recombinant(rTdT) 93
TES,Ultrapure 245
Tetracycline,Hydrochloride,Ultrapure 245
TetramethylammoniumChloride,Ultrapure 245
3,3’,5,5’-Tetramethylbenzidine 245
N,N,N’,N’-Tetramethylethylenediamine,Ultrapure 245
TetrazoliumRed—See2,3,5-TriphenylTetrazoliumChloride 246
Product Name Index | Alphabetical
289For bulk or alternate pack sizes, email us at [email protected].
Product Page Product Page
TFActivationELISAKits
AP-1ColdProbe 110
AP-1ELISAKit 110
AP-1ELISAKitwithNuclearExtractionKit 110
ControlHEK292PMANuclearExtract 110
E2F-1ELISAKit 110
E2F-1ELISAKitwithNuclearExtractionKit 110
ERaELISAKit 110
ERaELISAKitwithNuclearExtractionKit 110
EREColdProbe 110
ERControlMCF7NuclearExtract 110
FKHRELISAKit 110
FKHRELISAKitwithNuclearExtractionKit 110
GATA-1ColdProbe 110
GATA-1ControlRecombinantProtein 110
GATA-1ELISAKit 110
GATA-1ELISAKitwithNuclearExtractionKit 110
GRColdControlProbe 110
GRControlHela(DEX)NuclearExtract 110
GRELISAKit 110
GRELISAKitwithNuclearExtractionKit 110
HIFColdControlProbe 110
HIFControlCos1NuclearExtract 110
HIF-1aELISAKit 110
HIF-1aELISAKitwithNuclearExtractionKit 110
HNF-3bELISAKit 110
HNF-3bELISAKitwithNuclearExtractionKit 110
MEF-2ColdControlProbe 110
MEF-2ControlNuclearExtract 110
MEF-2ELISAKit 110
MEF-2ELISAKitwithNuclearExtractionKit 110
NFkBc-RelELISAKit 110
NFkBc-RelELISAKitwithNuclearExtractionKit 110
NFkBColdControlProbe 110
NFkBControlNuclearExtract 110
NFkBp50ELISAKit 110
NFkBp50ELISAKitwithNuclearExtractionKit 110
NFkBp52ELISAKit 110
NFkBp52ELISAKitwithNuclearExtractionKit 110
NFkBp65ELISAKit 110
NFkBp65ELISAKitwithNuclearExtractionKit 110
NFkBRel-bELISAKit 110
NFkBRel-bELISAKitwithNuclearExtractionKit 110
p53ColdControlProbe 110
p53ELISAKit 110
p53ELISAKitwithNuclearExtractionKit 110
PPARColdControlProbe 110
PPARaColdControlProbe 110
PPARaControlCos1NuclearExtract 110
PPARaELISAKit 110
PPARaELISAKitwithNuclearExtractionKit 110
SMAD-2ELISAKit 110
SMAD-2ELISAKitwithNuclearExtractionKit 110
SMAD-4ELISAKit 110
SMAD-4ELISAKitwithNuclearExtractionKit 110
SP-1ColdProbe 110
SP-1ControlRecombinantProtein 110
SP-1ELISAKit 110
SP-1ELISAKitwithNuclearExtractionKit 110
STAT-1ColdControlProbe 110
STAT-1ControlMCF7(IFNg) 110
STAT-1ELISAKit 110
STAT-1ELISAKitwithNuclearExtractionKit 110
TF-TFInteractionArrayI 115
TF-TFInteractionArrayII 115
TF-TFInteractionArrayIII 115
TF-TFInteractionIRefill 115
TF-TFInteractionIIRefill 115
TF-TFInteractionIIIRefill 115
TFF1 25
THBS2GenePromoterReporterVector 111
ThermoSequenaseCycleSequencingKit 172
ThermoSequenaseDyePrimerManualCycleSequencingKit 173
ThermoSequenaseKits
ThermoSequenaseCycleSequencingKit 172
ThermoSequenaseDyePrimerManualCycleSequencingKit 173
Thiamine,Hydrochloride 245
ThiaminePyrophosphateChloride—SeeCocarboxylase 207
Product Name Index | Alphabetical
290 888-362-2447 | 216-765-5000 | usb.affymetrix.com
Product Page Product Page
ThiazolylBlue—SeeMTT 228
Thimerosal 246
Thioredoxin,Human,Recombinant 246
L-Threonine 246
Thrombin 246
ThrombinHuman 246
TIMP3GenePromoterReporterVector 111
TMAC—SeeTetramethylammoniumChloride 245
TMB—See3,3’,5,5’-Tetramethylbenzidine 245
TNFaGenePromoterReporterVector 111
TopoisomeraseII,Alpha 95
ToxillicAcid—SeeMaleicAcid 226
TranscriptionFactors
EMSA(Gel-Shift)KitsandProbeSets 103
Function-SpecificProtein/DNAArrays 115
GenePromoterReporterVectors 111
LuciferaseReporterVectors 112
MammalianFull-lengthTFExpressionVectors 114
NFkB-TFandPPAR-TFInteractionArrays 118
Protein/DNAArrays 116
TFActivationELISAKits 107
TF-TFInteractionArrays 115
Transfection
DeliverXandDeliverXPlussiRNATransfectionKits 98
a,a-Trehalose,Dihydrate 246
Tricine,Ultrapure 246
Triethanolamine,Hydrochloride 246
1,3,7-Trimethylxanthine—SeeCaffeine 204
2,6,8-Trioxypurine—SeeUricAcid 249
2,3,5-TriphenylTetrazoliumChloride 246
TriphosphopyridineNucleotideDinucleotide—SeeNicotinamideAdenineDinucleotideReduced 230
2,4,6-Tripyridyl-s-Triazine(TPTZ) 247
Tris 247
Tris,Ultrapure 247
Tris-Glycine(TG)Buffer,10XSolution,Ultrapure 247
Tris-Glycine-SDS(TGS)Buffer,10XSolution,Ultrapure 247
Tris,Hydrochloride,Ultrapure 247
Tris/HCl,1MSolution,pH7.0,Ultrapure 248
Tris/HCl,1MSolution,pH7.5,Ultrapure 248
Tris/HCl,1MSolution,pH8.0,Ultrapure 248
r-hTRX—SeeThioredoxin,Human,Recombinant 246
Trypsin,Ultrapure 248
Trypsin1:250 248
Trypsin(TPCKTreated) 248
L-Tryptophan 248
TthDNAPolymerase 89
Two-StepRT-PCRKit 48
Tyrosinase 248
L-Tyrosine 248
U
Uracil 249
Uracil-DNAGlycosylase,E. coli 67
Uracil-DNAGlycosylase,Heat-Labile 67
Urea 249
Urea,Ultrapure 249
UricAcid 249
Uridine 249
Uridine-5’-Triphosphate,TrisodiumSalt 249
USF1CMVExpressionVector 114
UTP—SeeUridine-5’-Triphosphate 249
V
Vancomycin,Hydrochloride 250
VDRLuciferaseReporterVector 112
VeriQuestFastProbeqPCRMasterMix(2X) 36
VeriQuestFastProbeqPCRMasterMix,NoReferenceDye(2X) 37
VeriQuestFastSYBRGreenqPCRMasterMix(2X) 30
VeriQuestFastSYBRGreenqPCRMasterMixwithFluorescein(2X) 31
VeriQuestProbeOne-StepqRT-PCRMasterMix(2X) 42
VeriQuestProbeOne-StepqRT-PCRMasterMix,NoReferenceDye(2X) 43
VeriQuestProbeqPCRMasterMix(2X) 33
VeriQuestProbeqPCRMasterMix,NoReferenceDye(2X) 35
VeriQuestSYBRGreenOne-StepqRT-PCRMasterMix(2X) 40
VeriQuestSYBRGreenOne-StepqRT-PCRMasterMixwithFluorescein(2X) 41
VeriQuestSYBRGreenqPCRMasterMix(2X) 27
VeriQuestSYBRGreenqPCRMasterMixwithFluorescein(2X) 29
VeriQuestTaqDNAPolymerase 26
VHL 25
Product Name Index | Alphabetical
291For bulk or alternate pack sizes, email us at [email protected].
Product Page Product Page
VHLGenePromoterReporterVector 111
VitafreeCasein—SeeCaseinVitafree 205
VitaminB1—SeeThiamineHCl 245
VitaminB2—SeeRiboflavin 238
VitaminB5—SeeD-CalciumPantothenate 204
VitaminB12 250
VitaminC—SeeL-AscorbicAcid 201
VitaminFreeCasein—SeeCasein-Vitafree 205
VitaminG—SeeRiboflavin 238
VitaminK1 250
v-mycGenePromoterReporterVector 111
W
Water,Nuclease-Free,Ultrapure 250
Water,RNase-Free,DEPCTreated,Ultrapure 250
Wright’sStain 250
X
XanthineMonosodiumSalt,Monohydrate 250
X-a-GAL—See5-Bromo-4-Chloro-3-Indolyl-a-D-Galactopyranoside 203
X-Gal—See5-Bromo-4-Chloro-3-Indolyl-b-D-Galactopyranoside 203
XyleneCyanolFF,Ultrapure 251
D-(+)-Xylose 251
Y, Z
Yeast,Bakers 251
Yeast,Brewers 251
YeastExtractPowder,LowDust 251
YeastExtractPowder,Ultrapure 251
YeastHydrolysate,Enzymatic,Ultrapure 251
Yeast-Torula 251
2XYTBroth 251
YY1LuciferaseReporterVector 112
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CATALOG ISSUE 1.14