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PCR

PCR. Presented by : Mostafa al- mosavi. 1993 Nobel Prize in Chemistry. 1- Definition of PCR. 2- Requirements for PCR. 3-PCR Process. 4-Procedure. Definition of PCR.

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PCR

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  1. 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. Thermusaquaticus 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. Procedure

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

  11. 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.

  12. 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.

  13. 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.

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