Caister Academic Press

Mutation Detection by Real-Time PCR

Authors: K.J. Edwards and J.M.J Logan
Abstract: Real-time PCR is ideally suited for analysis of single nucleotide polymorphisms (SNPs) and has been increasingly used for this purpose since the advent of real-time PCR and as whole genome sequences have become available. It requires methods that are rapid, sensitive, specific and inexpensive, and several real-time methods have evolved which fulfil these requirements. Additionally real-time PCR is a technique that is readily amenable to automation and no post-PCR handling is required. Different formats have been applied including hybridisation probes with melting curve analysis, hydrolysis probes, molecular beacons and scorpion primers. SNP detection by real-time PCR has found applications in diagnosis of human disease, pharmacogenetics, clinical microbiology and drug development, and has replaced techniques such as sequencing, single strand conformation polymorphism and restriction enzyme digestion.
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