Protocols and Databases in Life Science

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Home PCR Nested PCR

Nested PCR

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What is nested PCR? Nested PCR is a variation of the polymerase chain reaction (PCR), in that two pairs (instead of one pair) of PCR primers are used to amplify a fragment.

The first pair of PCR primers amplify a fragment similar to a standard PCR. However, a second pair of primers called nested primers (as they lie / are nested within the first fragment) bind inside the first PCR product fragment to allow amplification of a second PCR product which is shorter than the first one.

The advantage of nested PCR is that if the wrong PCR fragment was amplified, the probability is quite low that the region would be amplified a second time by the second set of primers. Thus, Nested PCR is a very specific PCR amplification.

PCR amplification of cDNA segments by 2 stage nested PCR

Standard Nested PCR Protocol
C. elegans Gene Knockout Project, Vancouver Lab