Nested PCR

2015-01-13

azim58 - Nested PCR


Nested polymerase chain reaction is a modification of polymerase chain
reaction intended to reduce non-specific binding in products due to the
amplification of unexpected primer binding sites.

Nested polymerase chain reaction involves two sets of primers, used in
two successive runs of polymerase chain reaction, the second set intended
to amplify a secondary target within the first run product.

People often perform nested PCR by doing pcr with one set of primers.
They then use some volume from this first pcr reaction in a second pcr
reaction. "The second-round PCR is performed with two new primers that
anneal to sequences internal to the product of the first-round PCR. In
this way, only specific first-round PCR products (and not nonspecific
products) will be amplified in the second round. Alternatively, it is
possible to perform semi-nested PCR by using one internal and one
first-round primer in the second PCR"


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some references for nested pcr

Osamu Ohara 1989 (PMID: 2788276)
amp;cd=4&cad=rja&ved=0CEYQFjAD&url=http%3A%2F%2Fwww.csub.edu%2F
~kszick_miranda%2FNested%2520PCR.doc&ei=Abn2UdLcF4e5igLgmoGQAw&usg=
AFQjCNGaINUGDvgm0S-6O-4rfabLFEqEjA&sig2=nMPhw5XLAuLiRKNEQX-z2w&bvm=
bv.49967636,d.cGE

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see also