Nested polymerase chain reaction

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

The amount of product from the PCR increases with the number of temperature cycles that the reaction is subjected to.

A commonly occurring problem is primers binding to incorrect regions of the DNA, giving unexpected products.

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.

The target DNA undergoes the first run of polymerase chain reaction with the first set of primers, shown in green.