Molecular markers
Polymerase chain reaction variations
Nested PCR

Author: Rodney E. Shackelford, D.O., Ph.D. (see Authors page)

Revised: 4 June 2018, last major update July 2010

Copyright: (c) 2008-2018,, Inc.

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

Table of Contents
Definition / general
Cite this page: Shackelford, R.E. Nested PCR. website. Accessed August 16th, 2018.
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