Molecular markers
Polymerase chain reaction variations
Nested PCR



Topic Completed: 1 June 2010

Revised: 15 February 2019

Copyright: 2008-2018, PathologyOutlines.com, Inc.

PubMed Search: Nested PCR[TI] pathology full text[sb]


Rodney E. Shackelford, D.O., Ph.D.
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Table of Contents
Definition / general
Cite this page: Shackelford R. Nested PCR. PathologyOutlines.com website. http://www.pathologyoutlines.com/topic/molecularpathnestedpcr.html. Accessed August 19th, 2019.
Definition / general
  • Nested PCR uses two sets of primers, one set is internal to or "nested within" with other set
  • The first or "outer" pair of primers amplifies the target sequence; then a dilution is made of the reaction and the internal or nested primers are used to continue the amplification
  • Alternatively, the technique can be "seminested" where one original PCR primer and one internal primer are used after the first amplification
  • Both techniques will generate an amplified sequence shorter than the first amplified sequence
  • If the first reaction is amplified nonspecifically, then it is unlikely that the internal amplification would also proceed
  • Thus, this technique increases the specificity and sensitivity of the PCR reaction
  • A drawback of this technique is that the addition of new primers after the first amplification round increases the chances of nonspecific contamination; many clinical labs avoid this technique for this reason
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