Polymerase chain reaction
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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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Polymerase chain reaction

POLYMERASE CHAIN REACTION

PCR


Features

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


Procedure

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


Procedure1

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


Problems

Problems

  • Contamination of sample

  • Any source of DNA will do (bacteria, technician…..)

  • Sterile conditions needed.

© 2010 Paul Billiet ODWS


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