dna rekombinan

Bioteknologi Pertanian

Lisnawita

Mikroba

Rekombinan

Mikroba rekombinan

DNA rekombinan

Metode perakitan

DNA (Deoxyribonucleic acid)

adalah sejenis asam nukleat yang tergolong biomolekul utama penyusun berat kering setiap organisme

peran DNA di dalam sebuah sel adalah sebagai materi genetik

DNA adalah pembawa informasi genetik di dalam suatu sel

Kumpulan teknik atau motode yang

digunakan untuk mengkombinasikan

gen-gen secara buatan

Proses rekombinasi

Hasil rekombinan

Teknologi DNA rekombinan


Rekombinasi antara molekul DNA dari organisme hidup yang berbeda fenomena umum di alam

Contoh Corynebacterium diphtheriae yang diinfeksi oleh virus szlig menghasilkan toksin yang bertanggung jawab terhadap gejala dipteri

Perubahan genetik pada bakteri yang disebabkan virus merupakan rekayasa genetik di alam





Duplikasi fenomena alam di laboratorium

Mengembangkan metode introduksi berbagai

informasi genetik ke dalam suatu organisme

Produksi bahan mahal yang tidak mungkin

dibuat secara tradisional





PLASMID

Kromoson ndash vs - Plasmid

Memotong DNA

Menggunakan enzim restriksi yaitu

enzim yang memotong dsDNA (double

stranded DNA) pada situs spesifik

Menyambung DNA

Tahapan DNA rekombinan


(Kloning gen)

Kloning DNA ke plasmid

Video biotek Steps in Cloning a Gene -

YouTube [360p]mp4

Video biotekRecombinant DNA copy2004

Demonstratives Inc - YouTube [360p]mp4

Seleksi bakteri pembawa DNA rekombinan

Performing the Spread Plate method I

1 Choose appropriate nutrient agar plate with the correct antibiotic (and X-gal) if visual screening

2 Using sterile technique transfer a loopful of bacteria from a culture tube onto plate (or 100l of bacterial culture using a pipette)

3 Dip glass ldquohockey stickrdquo in 70 ethanol Holding it DOWNWARDS flame until alcohol is burned off DO NOT put back into alcohol

4 Remove lid of petri dish With one hand rotate dish With other hand move hockey stick lightly over surface to spread the inoculum evenly

Performing the Spread Plate method II

70 EtOH

ldquoSpreaderrdquo or ldquoHockey Stickrdquo

Keep flame away from alcohol

Performing a Plate Streak I

1 Flame metal inoculating loop let cool momentarily

2-3 Using sterile technique transfer a loop of bacterial culture or single colony onto loop

4 With one hand remove lid of dish With other hand lightly brush the loop back and forth on one quadrant of the dish

Performing a Plate Streak II

4 Reflame metal inoculating loop let cool momentarily

567 Rotate petri dish 90deg Use 1st streak as inoculum for 2nd streak (only pass the loop through the 1st streak once) Repeat once more rotating dish 90deg and sterilizing loop again

8 Incubate on at 37degC

Plate Streak Method

This is an example of a good streak for isolation using the four corners method

This is not a great streak plate but it is serviceable as there are a few isolated colonies - would have been better if the loop had been flamed between each sector

This is an example of how NOT to streak for isolation Scribbling is not streaking and most likely will not result in isolated colonies

After Incubating the plate overnight at 37degC- individual

colonies of transformed bacteria

should be seen

Each team will pick two individual colonies

(clones) and streak on a new plate (single

colony purification) for next week

Bacterial colonies transformed with pUC18

blue colonies (contain non-recombinant DNA molecules)

White colonies (contain recombinant DNA molecules)

Video biotek dna cloning - YouTube

[360p]mp4

Manfaat teknologi DNA rekombinan

Dengan mengisolasi dan mempelajari masing-

masing gen akan diperoleh pengetahuan

tentang fungsi dan mekanisme kontrolnya

teknologi ini memungkinkan diperolehnya

produk gen tertentu dalam waktu lebih cepat

dan jumlah lebih besar daripada produksi

secara konvensional

Deteksi keberadaan DNA sisipan

1 Pre Denaturasi

2 Denaturasi

3 Annealing

4 Ekstensi

5 Post ekstensi (pemantapan)

Tahapan-tahapan PCR

PCR Animation Please click here

1st cycle 2nd cycle 3rd cycle

Process

Denature

Anneal Primer

Replicate

DNA

1 2 5 4 M 8

10 11 6 7

391 bp

M M M 7 6 5 4

A B

3

238 bp

9

Contoh hasil PCR

Hasil penentuan urutan nukleotida DNA

sisipan (DNA sekuensing)

GR1-Jatim 2 TTGTTGTACGTGCCGTACCTTGCGGCATGTCTGCGCTTGTGTGCTACGTCCGTGGCCGTG 60

GR3-Jatim 3 TTGTTGCACGTGCCGTACCTTGCGGCATGTCTGCGCTTGTGTGCTACGTCCGTGGCCGTG 60

GR2-Jatim 1 TTGTTGTACGTGCCGTACCTTGCGGCATGTTTGCGCTTGTGTGCTACGTCCGTGGCCGTG 60

GR1-Jatim 2 ATGAGACGACGTGTTAGGACCCGTGCCTGGCATTGGCACGTGGTTTAAGACTTGATGAGT 120



GR1-Jatim 2 GCCCGCAGGCACCGCCAGCTTTTTCCCATTTTTATTTATTTTTTATGCAATTCGATTGCT 180



GR1-Jatim 2 AAAATATTCTAGTCTTATCGGTGGATCACTCGGCTCGTGGATCGATGAAGATCGCTAGCC 240

GR3-Jatim 3 AAAATATTCTAGTCTTATCGGTGGATCACTCGGCTCSKGGATCGATGAAGATCGCTAGCC 240

GR2-Jatim 1 AAAATATTCTAGTCTTATCGGTGGATCACTCGGCTCGKGGATCGATGAARATCGCTAGCC 240

GR1-Jatim 2 TTCTGCA 247



Hasil pembacaan DNA sekuensing

Bidang Hukum Pelaku kejahatan dapat diidentifikasi dengan menggunakan analisis Sidik jari DNA

Misalnya Kasus perkosaan

02 Hybrid Seed

14 Food

18 Starch

37 Alcohol

58 Sweeteners

447 Animal FeedResidual

168 Exports

256 Ending Stocks (Buffer against a bad crop)

Avg Annual US Usage

Oil is extracted from the germ (embryo) for cooking Starch in building materials

or intravenous solutions the shell (hull) is used in animal

feed

Source National Geographic June 1993 p91-117

Mikroba

Rekombinan

Mikroba rekombinan

DNA rekombinan

Metode perakitan








Proses rekombinasi

Hasil rekombinan



















PLASMID


Memotong DNA




Menyambung DNA



(Kloning gen)



YouTube [360p]mp4










70 EtOH











Plate Streak Method






should be seen








[360p]mp4








secara konvensional


1 Pre Denaturasi

2 Denaturasi

3 Annealing

4 Ekstensi


Tahapan-tahapan PCR



Process

Denature

Anneal Primer

Replicate

DNA

1 2 5 4 M 8

10 11 6 7

391 bp

M M M 7 6 5 4

A B

3

238 bp

9

Contoh hasil PCR





















02 Hybrid Seed

14 Food

18 Starch

37 Alcohol

58 Sweeteners


168 Exports


Avg Annual US Usage



feed


Mikroba rekombinan

DNA rekombinan

Metode perakitan








Proses rekombinasi

Hasil rekombinan



















PLASMID


Memotong DNA




Menyambung DNA



(Kloning gen)



YouTube [360p]mp4










70 EtOH











Plate Streak Method






should be seen








[360p]mp4








secara konvensional


1 Pre Denaturasi

2 Denaturasi

3 Annealing

4 Ekstensi


Tahapan-tahapan PCR



Process

Denature

Anneal Primer

Replicate

DNA

1 2 5 4 M 8

10 11 6 7

391 bp

M M M 7 6 5 4

A B

3

238 bp

9

Contoh hasil PCR





















02 Hybrid Seed

14 Food

18 Starch

37 Alcohol

58 Sweeteners


168 Exports


Avg Annual US Usage



feed









Proses rekombinasi

Hasil rekombinan



















PLASMID


Memotong DNA




Menyambung DNA



(Kloning gen)



YouTube [360p]mp4










70 EtOH











Plate Streak Method






should be seen








[360p]mp4








secara konvensional


1 Pre Denaturasi

2 Denaturasi

3 Annealing

4 Ekstensi


Tahapan-tahapan PCR



Process

Denature

Anneal Primer

Replicate

DNA

1 2 5 4 M 8

10 11 6 7

391 bp

M M M 7 6 5 4

A B

3

238 bp

9

Contoh hasil PCR





















02 Hybrid Seed

14 Food

18 Starch

37 Alcohol

58 Sweeteners


168 Exports


Avg Annual US Usage



feed



















PLASMID


Memotong DNA




Menyambung DNA



(Kloning gen)



YouTube [360p]mp4










70 EtOH











Plate Streak Method






should be seen








[360p]mp4








secara konvensional


1 Pre Denaturasi

2 Denaturasi

3 Annealing

4 Ekstensi


Tahapan-tahapan PCR



Process

Denature

Anneal Primer

Replicate

DNA

1 2 5 4 M 8

10 11 6 7

391 bp

M M M 7 6 5 4

A B

3

238 bp

9

Contoh hasil PCR





















02 Hybrid Seed

14 Food

18 Starch

37 Alcohol

58 Sweeteners


168 Exports


Avg Annual US Usage



feed



Memotong DNA




Menyambung DNA



(Kloning gen)



YouTube [360p]mp4










70 EtOH











Plate Streak Method






should be seen








[360p]mp4








secara konvensional


1 Pre Denaturasi

2 Denaturasi

3 Annealing

4 Ekstensi


Tahapan-tahapan PCR



Process

Denature

Anneal Primer

Replicate

DNA

1 2 5 4 M 8

10 11 6 7

391 bp

M M M 7 6 5 4

A B

3

238 bp

9

Contoh hasil PCR





















02 Hybrid Seed

14 Food

18 Starch

37 Alcohol

58 Sweeteners


168 Exports


Avg Annual US Usage



feed


Menyambung DNA



(Kloning gen)



YouTube [360p]mp4










70 EtOH











Plate Streak Method






should be seen








[360p]mp4








secara konvensional


1 Pre Denaturasi

2 Denaturasi

3 Annealing

4 Ekstensi


Tahapan-tahapan PCR



Process

Denature

Anneal Primer

Replicate

DNA

1 2 5 4 M 8

10 11 6 7

391 bp

M M M 7 6 5 4

A B

3

238 bp

9

Contoh hasil PCR





















02 Hybrid Seed

14 Food

18 Starch

37 Alcohol

58 Sweeteners


168 Exports


Avg Annual US Usage



feed




(Kloning gen)



YouTube [360p]mp4










70 EtOH











Plate Streak Method






should be seen








[360p]mp4








secara konvensional


1 Pre Denaturasi

2 Denaturasi

3 Annealing

4 Ekstensi


Tahapan-tahapan PCR



Process

Denature

Anneal Primer

Replicate

DNA

1 2 5 4 M 8

10 11 6 7

391 bp

M M M 7 6 5 4

A B

3

238 bp

9

Contoh hasil PCR





















02 Hybrid Seed

14 Food

18 Starch

37 Alcohol

58 Sweeteners


168 Exports


Avg Annual US Usage



feed




YouTube [360p]mp4










70 EtOH











Plate Streak Method






should be seen








[360p]mp4








secara konvensional


1 Pre Denaturasi

2 Denaturasi

3 Annealing

4 Ekstensi


Tahapan-tahapan PCR



Process

Denature

Anneal Primer

Replicate

DNA

1 2 5 4 M 8

10 11 6 7

391 bp

M M M 7 6 5 4

A B

3

238 bp

9

Contoh hasil PCR





















02 Hybrid Seed

14 Food

18 Starch

37 Alcohol

58 Sweeteners


168 Exports


Avg Annual US Usage



feed











70 EtOH











Plate Streak Method






should be seen








[360p]mp4








secara konvensional


1 Pre Denaturasi

2 Denaturasi

3 Annealing

4 Ekstensi


Tahapan-tahapan PCR



Process

Denature

Anneal Primer

Replicate

DNA

1 2 5 4 M 8

10 11 6 7

391 bp

M M M 7 6 5 4

A B

3

238 bp

9

Contoh hasil PCR





















02 Hybrid Seed

14 Food

18 Starch

37 Alcohol

58 Sweeteners


168 Exports


Avg Annual US Usage



feed










Plate Streak Method






should be seen








[360p]mp4








secara konvensional


1 Pre Denaturasi

2 Denaturasi

3 Annealing

4 Ekstensi


Tahapan-tahapan PCR



Process

Denature

Anneal Primer

Replicate

DNA

1 2 5 4 M 8

10 11 6 7

391 bp

M M M 7 6 5 4

A B

3

238 bp

9

Contoh hasil PCR





















02 Hybrid Seed

14 Food

18 Starch

37 Alcohol

58 Sweeteners


168 Exports


Avg Annual US Usage



feed






should be seen








[360p]mp4








secara konvensional


1 Pre Denaturasi

2 Denaturasi

3 Annealing

4 Ekstensi


Tahapan-tahapan PCR



Process

Denature

Anneal Primer

Replicate

DNA

1 2 5 4 M 8

10 11 6 7

391 bp

M M M 7 6 5 4

A B

3

238 bp

9

Contoh hasil PCR





















02 Hybrid Seed

14 Food

18 Starch

37 Alcohol

58 Sweeteners


168 Exports


Avg Annual US Usage



feed






[360p]mp4








secara konvensional


1 Pre Denaturasi

2 Denaturasi

3 Annealing

4 Ekstensi


Tahapan-tahapan PCR



Process

Denature

Anneal Primer

Replicate

DNA

1 2 5 4 M 8

10 11 6 7

391 bp

M M M 7 6 5 4

A B

3

238 bp

9

Contoh hasil PCR





















02 Hybrid Seed

14 Food

18 Starch

37 Alcohol

58 Sweeteners


168 Exports


Avg Annual US Usage



feed









secara konvensional


1 Pre Denaturasi

2 Denaturasi

3 Annealing

4 Ekstensi


Tahapan-tahapan PCR



Process

Denature

Anneal Primer

Replicate

DNA

1 2 5 4 M 8

10 11 6 7

391 bp

M M M 7 6 5 4

A B

3

238 bp

9

Contoh hasil PCR





















02 Hybrid Seed

14 Food

18 Starch

37 Alcohol

58 Sweeteners


168 Exports


Avg Annual US Usage



feed



1 Pre Denaturasi

2 Denaturasi

3 Annealing

4 Ekstensi


Tahapan-tahapan PCR



Process

Denature

Anneal Primer

Replicate

DNA

1 2 5 4 M 8

10 11 6 7

391 bp

M M M 7 6 5 4

A B

3

238 bp

9

Contoh hasil PCR





















02 Hybrid Seed

14 Food

18 Starch

37 Alcohol

58 Sweeteners


168 Exports


Avg Annual US Usage



feed




Process

Denature

Anneal Primer

Replicate

DNA

1 2 5 4 M 8

10 11 6 7

391 bp

M M M 7 6 5 4

A B

3

238 bp

9

Contoh hasil PCR





















02 Hybrid Seed

14 Food

18 Starch

37 Alcohol

58 Sweeteners


168 Exports


Avg Annual US Usage



feed


1 2 5 4 M 8

10 11 6 7

391 bp

M M M 7 6 5 4

A B

3

238 bp

9

Contoh hasil PCR





















02 Hybrid Seed

14 Food

18 Starch

37 Alcohol

58 Sweeteners


168 Exports


Avg Annual US Usage



feed






















02 Hybrid Seed

14 Food

18 Starch

37 Alcohol

58 Sweeteners


168 Exports


Avg Annual US Usage



feed


dna rekombinan

Education

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