video on YouTube https://www.youtube.com/watch?v=NLmonT3GUHE https://www.youtube.com/watch?v=NLmonT3GUHE Polymerase Chain Reaction, PCR  is a method widely used in molecular biology to rapidly make millions to billions of copies of a specific DNA sample allowing scientists to take a very small sample of DNA and amplify it to a large enough amount to study in detail.

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PCR
Ahmed
Polymerase Chain Reaction

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PCR
is a method widely used in molecular biology to rapidly make millions
to billions of copies of a specific DNA sample allowing scientists to
take a very small sample of DNA and amplify it to a large enough
amount to study in detail.
Polymerase Chain Reaction
Kary Mullis94 c° 40 c° 72 c°

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PCR
What is components required in PCR
2. DNA polymerase
Taq Polymerase
1. Template (DNA)
Thermus aquaticus
3. Mg++ (MgCl2)
half-life of 40 min at 95°C
60 bases per sec at 70 °C
can amplify lengths of about 5 kb
catalyze phosphodiester bond formation
Facilitates formation primers and
DNA templates complex
by stabilizing negative charges on
their phosphate backbones

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PCR
4. Primers (Forward and Reverse)
Not pairing with each other
15–30 bases
Complementary
To strands
Similar melting
temperatures
5. Deoxynucleoside triphosphates
6. Buffer
Binding sites must be unique
GC content 40–60%
dGTP dATP dTTPdCTP
Suitable chemical environment for activity of DNA polymerase.
PH 8.0 and 9.5

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PCR
PCR Cycle
Denaturation
95
Annealing
45-65
Extension
72
Denaturation
Annealing
Extension
Unwinding the double helix by heating to 95 degrees
Priming the DNA by cooling the test tube to 45-65 degrees
Adding on complementary nucleotides and reheating to 72
degrees

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PCRDenaturation 95°CAnnealing 60°CExtension 72°C