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Transcript of Primer Design.ppt
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PRIMER DESIGN
Selasa, 5 April 2016
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PRIMER ?
A pair of synthetic oligonucleotides to prime
DNA synthesis (Sambrook & Russell, 200!
"eniru primer RNA yang menga#ali prosessintesis DNA (replikasi! di dalam sel$
Dipakai pada teknik %R dan sekuensing DNA
(menentukan keberhasilan teknik ini!
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PRIMER ?
%rimer di'design memiliki urutan nukleotida
yang komplemen terhadap sekuen DNAcetakan (template! sehingga dapat menempel
dengan baik$
3-AGCTGTACGGCATGCGTAGCCTTATGATGC-5 (DA !e"a#a$%
5-TCGACATGCCGTACGCATC-3 (Pri&er%
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'""p))r*sali$+i$*)
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.EAPA PER/ DESAI
PRIMER??
Is reire+ "* *"ai$ "'e +esire+ (PCR% pr*+!"s
i$ 'i4' iel+, "* sppress a&plii!a"i*$ *
$a$"e+ see$!es, a$+ "* a!ili"a"essee$" &a$ipla"i*$ * "'e a&pliie+
pr*+!" (Sa&r**# 7 Rssell, 2001%
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.AIDA8 DA/AM DESAI
PRIMER??
Pa$9a$4 pri&er
"elting temperature(T&%
Specificity (#e$i#a$ DA "ar4e"%
r"a$ asa-
G)C !*$"e$" +a$ seara$ p*lpri&i+i$ (T, C%
+a$ p*lpri$e (A, G%
r"a$ asa pa+a 9$4 3:(;i$*+, 200
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PA=AG PRIMER??
Me$e$"#a$ specificity target DNA
Se&a#i$ pa$9a$4, se&a#i$ spesii#
Terlal pe$+e#, &+a' &e$e&pel)"i+a# spesii#
>er#*relasi +e$4a$ eisie$si annealing
Se&a#i$ pa$9a$4, se&a#i$ eisie$
Pa$9a$4 pri&er er#isar 1-2< asa-
Pa$9a$4 pri&er "er#ai" A+a$ &
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"elting emperature (&%
S' saa" se"e$4a' +ari i#a"a$ ra$4#ap DA
&e$9a+i "$44al
&+ari sepasa$4 pri&er +i+esai$ &e&ili#i $ilai
a$4 sa&a, 9i#a ere+a &a#a si$"esis
(a&plii#asi% DA "i+a# *p"i&al pri&er +4$ &"i$44i "i+a# isa &e$e&pel pa+a s' re$+a',
se+a$4#a$ pri&er +4$ &re$+a' "i+a# isa
&e$e&pel pa+a s' "i$44i
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& Dapa" +i'i"$4 +e$4a$ r&s)
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Se!ara &&, &a$4 &e$4'asil#a$ pr*+#
PCR a$4 ai# a$"ara 55-@20C (sesai +e$4a$r&s alla!e $"# pa$9a$4 pri&er 1-2ERAPA I/AI GC !*$"e$"-$a?
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Te$"#a$ $ilai T& #a$+i+a" pri&er (*rar+ 7
reerse%, &isal$a +e$4a$ &e"*+e alla!e (I$4a",pere+aa$ $ilai T& #e+a$a &a#s 50C%
5-ATGGTTTCCAGAGAATGGTG-3
>ERAPA I/AI T&-$a?
5-TCATCTTGAAAGCAATTGAG -3
>ERAPA I/AI T&-$a?
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Ce# #e&$4#i$a$ "er9a+i$a sel-+i&er a"a
'airpi$ l**p se!ara &a$al a"a &e$44$a#a$pera$4#a" l$a#, &isal$a PearlPri&er (+apa" +i-
+*$l*a+ +i '""p))perlpri&ers*r!e*r4e$e" )%
http://perlprimer.sourceforge.net/http://perlprimer.sourceforge.net/http://perlprimer.sourceforge.net/http://perlprimer.sourceforge.net/ -
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Desain Primer denan
Sot'are/(n)line
Me$44$a#a$ pera$4#a" l$a# PearlPri&er
Se!ara *$li$e '""p))si&4e$e!*&)Pri&er3,
'""p))$!i$l&$i'4*)"**ls)pri&er-las"),
li4*@, +ll
http://simgene.com/Primer3http://www.ncbi.nlm.nih.gov/tools/primer-blast/http://www.ncbi.nlm.nih.gov/tools/primer-blast/http://simgene.com/Primer3 -
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*+#I,+N
Desai$la' pri&er $"# "ar4e" 4e$ eri#"
4iF556@@5F4F.C@H
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Thank You
KingsoftOceMake Presentation much more fun
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per"a$aa$
Gi&a$a !ara$a $"# &e$e$"#a$
a$a#$a pri&er rerse+a$ *rar+ saa"
+esai$ pri&er ?
