PCR – Polymerase Chain Reaction A method of amplifying small amounts of DNA using the principles of DNA replication.

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Presentation transcript:

PCR – Polymerase Chain Reaction A method of amplifying small amounts of DNA using the principles of DNA replication

DNA Replication Normal DNA replication requires the following: DNA gyrase DNA helicase SSBs DNA polymerase III andI DNA ligase RNA primers and primase Many nucleotides and template DNA

Polymerase Chain Reaction PCR only requires: Single strand DNA fragment Polymerase Primer Heat provides the mechanism for H bonds to release, replacing gyrase, helicase, SSBs and primase DNA primers replace RNA primers

PCR Machine

Taq Polymerase Taq polymerases comes from a bacteria that lives in themrmal hotsprings and can tolerate the high heat of PCR (9r degrees C) These bacteria were discovered in 1966, living in Yellowstone hot springs The heat tolerant bacteria is essential to PCR

PCR Process DNA sample is heated to 95 degrees DNA denatures, H bonds break Sample is cooled to 60 degrees DNA primers anneal to complementary sections Taq polymerase enzymes build a complementary strand DNA sample is heated to 95 degrees…

…2 hours later… In 32 cycles at 100% efficiency, 1.07 billion copies of targeted DNA region are created. 20 cycles takes about 1 hour Amplified sample can be used in DNA fingerprinting, sequencing, recombinant DNA, etc…

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