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POLYMERASE CHAIN REACTION

POLYMERASE CHAIN REACTION. PCR. Image Credit: Taq DNA polymerase . 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 few hours.

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POLYMERASE CHAIN REACTION

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  1. POLYMERASE CHAIN REACTION PCR

  2. Image Credit: Taq DNA polymerase 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 few hours © 2010 Paul Billiet ODWS

  3. Procedure • Mix: DNA sample, specific complementary primers, DNA polymerase • Heat DNA to 95°C, denatures DNASister strands separate • Cool to 56°C Primers bind to DNABracket region to be copiedStop sister strand reannealing © 2010 Paul Billiet ODWS

  4. Procedure • Warm to 73°COptimum temperature for DNA polymeraseNew sister strands synthesised from the primers • Heat again to 95°CStrands 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 © 2010 Paul Billiet ODWS

  5. Problems • Contamination of sample • Any source of DNA will do (bacteria, technician…..) • Sterile conditions needed. © 2010 Paul Billiet ODWS

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