Novel amplification of DNA in a hairpin structure: towards a radical elimination of PCR errors from amplified DNA.

ABSTRACT: Errors introduced during PCR amplification set a selectivity limit for microsatellite analysis and molecular mutation detection methods since polymerase misincorporations invariably get confused with genuine mutations. Here we present hairpin-PCR, a new form of PCR that completely separates genuine mutations from polymerase misincorporations. Hairpin-PCR operates by converting a DNA sequence to a hairpin following ligation of oligonucleotide caps to DNA ends. We developed conditions that allow a DNA hairpin to be efficiently PCR-amplified so that, during DNA synthesis, the polymerase copies both DNA strands in a single pass. Consequently, when a misincorporation occurs it forms a mismatch following DNA amplification, and is distinguished from genuine mutations that remain fully matched. Er

SUBMITTER: Kaur M 

PROVIDER: S-EPMC152880 | biostudies-literature | 2003 Mar

REPOSITORIES: biostudies-literature