Biology Protocols

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What is PCR?

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What is PCR?

PCR refers to Polymerase Chain Reaction, a technique whereby DNA is amplified in vitro using an enzyme (Polymerase) in a series of temperature dependent steps, multiple times using short oligo primers, dNTPs and the correct buffer.

The step involved are common between most PCR protocols and incorporate a Denaturation step (94˚C for ~1 minute to denature the DNA strans of the template), Annealing Step (~5˚C below the melting temperature (Tm) of the DNA primers used for 30 seconds to allow annealing of the oligo primers to the DNA template), and Elongation (~68˚C for 30 secs per Kbp of DNA template, dependent on Polymerase used).

Various DNA polymerases are available for use in PCR, including general polymerases such as Taq, polymerases for longer templates such as Elongase (Invitrogen) and mnay others, each with varying degrees of speed, error (proof-reading) and action (overhangs, modified nucleotide incorporation etc.).


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