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Polymerase Chain Reaction

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Western University
Biochemistry 2288A
Chris Brandl

PCRAnother way to get a lot of your gene You can start with one copy of dna and end up with a lot Powerful way to get lot of your favorite genecloning is time consuming PCR takes only couple hours howevercloning takes weeksExponential amplification of any DNA from a source in which it is found as little as ONCE Extremely sensitiveInvolves the repeated extension of two primers that flank YFGPCR is replication reaction Key repeated multiple timesReagents 1Template DNA 2Two oligonucelotide primers which flank YGF usually one up stream and one down stream 3dNTPs since its replication reaction 4DNA polymerase 3 principle steps repeated over and overWe got template dnafirst step heat to denature strand 95degrees this separates two strandsSecond step hybridization reaction where you use oligo nucleotide to flank your geneone it red and one in greendo that around 5055 degrees primers will anneal to one strand and the other Third step dna synthesis in presence of dna polymerase and dntpwhere you start with one dna you got 2 dnas Extension reaction is done at
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