1. Presented by : Mostafa al-mosavi
PCR
Presented by :
Mostafa al-mosavi

2. 1993 Nobel Prize in Chemistry

3. 1- Definition of PCR.
2- Requirements for PCR.
3-PCR Process.
4-Procedure.

4. Definition of PCR
Polymerase chain reaction (PCR) enables researchers to produce millions of copies of a specific DNA sequence in two hours.

5. Requirements for PCR 1- DNA sample :
Very small amount (ng or some times less).
2- Two primers:
(Forward)a.
b. (Reverse)
You must know the sequence of the flaking regions so you can order appropriate primers.

6. Thermus aquaticus
3- Heat stable polymerase.
The bacterium Thermus aquaticus was first discovered in several springs in the Great Fountain area of the Lower Geyser Basin at Yellowstone National Park

7. 4-Four dNTPs.(Deoxy nuleoside tri phosphates)
Nitrogen Base dNTPs
Adenine
Thymine
Guanine
Cytosine

8. Requirements for PCR
5- Buffer(10x).mgcl2
6- DDW

9. Hot Star Tag Polymerase
Procedure
Volume(µl)
Reagents
2.5
10x PCR buffer 2
dNTPs
0.6
Forward Primer
Reverse Primer
0.3
Hot Star Tag Polymerase 17
Distilled water
DNA template 25
Total volume

10. 7-Thermocycler:
Change temperature very rapidly for each cycle.

13. The cycling reactions
There are three major steps in a PCR, which are repeated for 30 or 40 cycles. This is done on an automated cycler, which can heat and cool the tubes with the reaction mixture in a very short time.

14. PCR Process PCR –reaction is divided into 3 steps:
1-Denatration: During denatration,
the template DNA is seprated into its 2 separate by heating at the temperature about 95ºC.

15. PCR Process 2-Anneling:
This involves the annealing of the primer to the denatured.
3-Extension:
The synthesizing ,take place at a temperature of around 72ºC,this corresponds to the optimal temperature for the Tag-polymerase to work.