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POLYMERASE CHAIN REACTION

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Presentation on theme: "POLYMERASE CHAIN REACTION"— Presentation transcript:

1 POLYMERASE CHAIN REACTION
PCR © 2016 Paul Billiet ODWS

2 Features Amplifies small samples of DNA into millions of copies
Can copy the DNA of a single cell Uses DNA polymerase from hot spring bacteria (Thermus aquaticus) Automated to produce millions of copies in a few hours. Taq DNA polymerase © 2016 Paul Billiet ODWS

3 Procedure Mix: DNA sample, specific complementary primers, DNA polymerase Heat DNA to 95°C, denatures DNA Sister strands separate Cool to 56°C Primers bind to DNA Bracket region to be copied Stop sister strand reannealing. © 2016 Paul Billiet ODWS

4 Procedure Warm to 73°C Optimum temperature for DNA polymerase New sister strands synthesised from the primers Heat again to 95°C Strands separate again Cool to 56°C more primers bind Warm to 73°C DNA polymerase synthesises new sister strands Repeat cycle 10 times > 1000 copies Repeat cycle 20 times > 1 million copies. © 2016 Paul Billiet ODWS

5 Problems Contamination of sample
Any source of DNA will do (bacteria, technician…..) Sterile conditions needed. © 2016 Paul Billiet ODWS

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