Slide 1

: PCR PCR 1985 - 1989 1989 1985

5- X . RFLP: :- PCR :1- DNA.3- . . RFLP(Restriction Fragment Length Polymorphism) 2- Restriction of DNA .4- Screening by probe. RFLP:

- endonucleases DNA .1- DNA :2- Restriction of DNA : .- DNA DNA .

- ( Eco RI, RII) 4 6 bp Eco RI E. coli 6 bases .5\ .G / AATTC.3\ 5\ ..... GAATTC.3\ 109 : 1011 bp Base pair (bp) : (A=T, GC).

Kilo base pair = 1000 bp Mega base pair = 1000,000 bp 109 = 1000 Mbp 1011 = 100000 Mbp DNA . .

3- : electrophoresis analysis .4- Screening by probe :Probe: DNA ( RNA) sequence DNA ( RNA) probe DNA Plate. Screening by probe .

5- X : X bands .1- . RFLP:4- DNA.2- .3- .

: PCR:- PCR Dr. Kary Mullis, 1985 1993 . PCR- DNA DNA . Polymerase Chain Reaction (PCR) : Polymerase.- DNA .

PCR:5- MgCl2 .1- Template DNA. 2- Two oligonucleotide primers. 4- Taq polymerase. 3- dNTPs.

1- Template DNA: DNA "Template DNA".2- Two oligonucleotide primers: Primer: DNA ( RNA) DNA DNA OH- 3\ DNA .

3- dNTPs :dNTPs : .4- Taq polymerase: DNA pol. I E. coli PCR (72 ).5- MgCl2 . Taq pol. Thermus aquaticus .

PCR:1- Denaturation (92 95) 94C 1 min . 2- Annealing 37C 30 sec. 3- Extention 68 - 72C 1 min. 4- Cycling 30 35 cyc..

1- Denaturation (92 95) 94C 1 min : DNA DNA 94 .2- Annealing 37C 30 sec: primer DNA Template . 37 30 30 60 : - - G/C , A/T".

3- Extention 68 - 72C 1 min: Taq pol. primer primer Template 72 .4- Cycling 30 35 cyc.: 30 35 2n DNA n = .

electrophoresis analysis .

PCR (RFLP):

5- .1- DNA .2- PCR (PCR techniques) .3- .4- PCR techniques DNA .