The document discusses recombinant DNA technology. It explains that recombinant DNA technology involves combining genetic material from different organisms artificially using molecular cloning techniques. This allows genes to be inserted into host organisms in order to mass produce valuable proteins. Specific techniques discussed include using restriction enzymes to cut DNA at specific sites, ligating DNA fragments together, cloning DNA into plasmids, transforming bacteria with recombinant plasmids, and selecting and identifying transformed bacterial colonies. The document notes applications of this technology include identifying criminal suspects using DNA fingerprinting and producing important compounds that are difficult to obtain through traditional methods.

1. Bioteknologi Pertanian
Lisnawita

2. Mikroba
Rekombinan

3.  Mikroba rekombinan
 DNA rekombinan
Metode perakitan

4. 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

5. Kumpulan teknik atau motode yang
digunakan untuk mengkombinasikan
gen-gen secara buatan
Proses : rekombinasi
Hasil : rekombinan
Teknologi DNA rekombinan

6. Teknologi DNA rekombinan
 Rekombinasi antara molekul DNA dari
organisme hidup yang berbeda : fenomena
umum di alam
 Contoh : Corynebacterium diphtheriae yang
diinfeksi oleh virus ß menghasilkan toksin
yang bertanggung jawab terhadap gejala
dipteri
 Perubahan genetik pada bakteri yang
disebabkan virus merupakan rekayasa genetik
di alam

7. Teknologi DNA rekombinan

8. Teknologi DNA rekombinan

9. Teknologi DNA rekombinan

10. Teknologi DNA rekombinan

11. Duplikasi fenomena alam di laboratorium
 Mengembangkan metode introduksi berbagai
informasi genetik ke dalam suatu organisme
 Produksi bahan mahal yang tidak mungkin
dibuat secara tradisional
Teknologi DNA rekombinan

12. Teknologi DNA rekombinan

13. Teknologi DNA rekombinan

14. Teknologi DNA rekombinan
PLASMID :

15. Kromoson – vs - Plasmid

16. Memotong DNA
Menggunakan enzim restriksi yaitu :
enzim yang memotong dsDNA (double
stranded DNA) pada situs spesifik.

19. Menyambung DNA

20. Tahapan DNA rekombinan

21. Teknologi DNA rekombinan
(Kloning gen)

22. Kloning DNA ke plasmid

23.  ..Video biotek▶ Steps in Cloning a Gene -
YouTube [360p].mp4

24.  ..Video biotekRecombinant DNA ©2004
Demonstratives, Inc. - YouTube [360p].mp4

25. Seleksi bakteri pembawa
DNA rekombinan

27. 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 100l of bacterial
culture using a pipette)

28. 3: Dip glass “hockey stick” 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
“Spreader” or
“Hockey Stick”
Keep flame
away from
alcohol !!

29. 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

30. Performing a Plate Streak II
4: Reflame metal
inoculating loop,
let cool
momentarily.
5,6,7: Rotate petri dish 90° Use 1st streak as
inoculum for 2nd streak (only pass the loop
through the 1st streak once). Repeat once
more rotating dish 90° and sterilizing loop again
8 :Incubate o/n at 37°C

31. Plate Streak Method
This is an example of a good streak for isolation using the "four
corners" method.

32. 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.

33. After Incubating the
plate overnight at
37°C- 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

34. Bacterial colonies transformed with pUC18
blue colonies
(contain non-recombinant DNA
molecules)
White colonies
(contain recombinant DNA
molecules)

35.  ..Video biotek▶ dna cloning - YouTube
[360p].mp4

36. 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.

37. Deteksi keberadaan DNA sisipan

38. 1. Pre Denaturasi
2. Denaturasi
3. Annealing
4. Ekstensi
5. Post ekstensi (pemantapan)
Tahapan-tahapan PCR

39. PCR Animation
Please click here.
1st cycle 2nd cycle 3rd cycle
Process
Denature
Anneal Primer
Replicate
DNA

41. 1 2 54M 8
10
116 7
391 bp
M MM 7654
A B
3
238 bp
9
Contoh hasil PCR

42. Hasil penentuan urutan nukleotida DNA
sisipan (DNA sekuensing)

43. GR1-Jatim 2 TTGTTGTACGTGCCGTACCTTGCGGCATGTCTGCGCTTGTGTGCTACGTCCGTGGCCGTG 60
GR3-Jatim 3 TTGTTGCACGTGCCGTACCTTGCGGCATGTCTGCGCTTGTGTGCTACGTCCGTGGCCGTG 60
GR2-Jatim 1 TTGTTGTACGTGCCGTACCTTGCGGCATGTTTGCGCTTGTGTGCTACGTCCGTGGCCGTG 60
****** *********************** *****************************
GR1-Jatim 2 ATGAGACGACGTGTTAGGACCCGTGCCTGGCATTGGCACGTGGTTTAAGACTTGATGAGT 120
GR3-Jatim 3 ATGAGACGACGTGTTAGGACCCGTGCCTGGCATTGGCACGTGGTTTAAGACTTGATGAGT 120
GR2-Jatim 1 ATGAGACGACGTGTTAGGACCCGTGCCTGGCATTGGCACGTGGTTTAAGACTTGATGAGT 120
************************************************************
GR1-Jatim 2 GCCCGCAGGCACCGCCAGCTTTTTCCCATTTTTATTTATTTTTTATGCAATTCGATTGCT 180
GR3-Jatim 3 GCCCGCAGGCACCGCCAGCTTTTTCCCATTTTTATTTATTTTTTATGCAATTCGATTGCT 180
GR2-Jatim 1 GCCCGCAGGCACCGCCAGCTTTTTCCCATTTTTATTTATTTTTTATGCAATTCGATTGCT 180
************************************************************
GR1-Jatim 2 AAAATATTCTAGTCTTATCGGTGGATCACTCGGCTCGTGGATCGATGAAGATCGCTAGCC 240
GR3-Jatim 3 AAAATATTCTAGTCTTATCGGTGGATCACTCGGCTCSKGGATCGATGAAGATCGCTAGCC 240
GR2-Jatim 1 AAAATATTCTAGTCTTATCGGTGGATCACTCGGCTCGKGGATCGATGAARATCGCTAGCC 240
************************************ *********** **********
GR1-Jatim 2 TTCTGCA 247
GR3-Jatim 3 TTCTGCA 247
GR2-Jatim 1 TTCTGCA 247
*******
Hasil pembacaan DNA sekuensing

45. Bidang Hukum
Pelaku kejahatan dapat diidentifikasi dengan
menggunakan analisis Sidik jari DNA .
Misalnya : Kasus perkosaan

49. 0.2% Hybrid Seed
1.4% Food
1.8% Starch
3.7% Alcohol
5.8% Sweeteners
44.7% Animal
Feed/Residual
16.8% Exports
25.6% 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