Caister Academic Press

An Introduction to Real-Time PCR

Authors: N. A. Saunders
Abstract: The development of instruments that allowed real-time monitoring of fluorescence within PCR reaction vessels was a very significant advance. The technology is very flexible and many alternative instruments and fluorescent probe systems have been developed and are currently available. Real-time PCR assays can be completed very rapidly since no manipulations are required post-amplification. Identification of the amplification products by probe detection in real- time is highly accurate compared with size analysis on gels. Analysis of the progress of the reaction allows accurate quantification of the target sequence over a very wide dynamic range, provided suitable standards are available. Further investigation of the real-time PCR products within the original reaction mixture using probes and melting analysis can detect sequence variants including single base mutations. Since the first practical demonstration of the concept real-time PCR has found applications in many branches of biological science. Applications include gene expression analysis, the diagnosis of infectious disease and human genetic testing.
An Introduction to Real-Time PCR: Download full text