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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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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