Mutation Detection by Real-Time PCR
Elaine Lyon, Rong Mao and Jeffrey Swensen
from: Real-Time PCR: Current Technology and Applications (Edited by: Julie Logan, Kirstin Edwards and Nick Saunders). Caister Academic Press, U.K. (2009)
Abstract
Real-time applications for mutation detection include detecting alterations associated with inherited disease, acquired alterations in oncology, and microbial or viral mutations associated with drug resistance in infectious diseases. Probe chemistries described for these applications include hydrolysis (TaqMan®) and hybridisation probes (FRET and Molecular Beacons). Hydrolysis probes detect mutations by allele specific hybridisation at a specific temperature, while hybridisation probes allow dynamic detection through a temperature range. Primer chemistries are described for allele specific amplification and Scorpion primers. Recent progress in scanning amplicons for mutations also includes high resolution melting. The design of each of these methods is described, along with applications read more ...
