Modern Medical Laboratory Journal

Nested polymerase chain reaction (Nested PCR) is a modification of PCR that was designed to improve sensitivity and specificity. This technique reduces nonspecific amplification of the DNA template. Nested PCR involves the use of two primer sets and two successive PCR reactions. The first set of primers are designed to anneal to sequences upstream from the second set of primers and are used in an initial PCR reaction. The first reaction is performed with primers that cover the target sequence and some additional sequence flanking both ends of the target sequence. After the first reaction, a second reaction is performed on the products of the first PCR with primers that bind to the target sequence and are within the amplified sequence of the first PCR. This reduces the amount of nonspecific binding because in the second reaction, most of the amplicons of the first reaction only contain the target sequence and its surrounding sequences.
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