Polymerase Chain Reaction (PCR) is a powerful method to specifically amplify stretches of DNA from the complex background of the genome. Nature Methods presents a strategy to increase the number of targets that are amplified in one reaction.
PCR requires two primers for every DNA sequence to be amplified, one on each end. Since these primer sequences are unique to the target region, in theory, primers for many different targets could be used simultaneously in the same PCR reaction, also known as a multiplex reaction. In practice the presence of multiple primers leads to unspecific priming and the formation of primer-dimers, both of which produce artifacts and severely limit the efficacy of the amplification.
To minimize these limitations Anthony Brookes and colleagues developed a solid-phase amplification method, where primers are physically separated from each other by immobilization on a solid surface. The authors show that this MegaPlex-PCR strategy can amplify 75 genomic regions selected at random. They also demonstrate that no complicated pre-selection of primers is needed, and primers against any sequence of choice yield high quality results.
MegaPlex PCR will be of interest to researchers needing to amplify many different regions, such as exons or promoters, from complex genomes in a single reaction.
Anthony Brookes (University of Leicester, UK)
Abstract available online.
(C) Nature Methods press release.
Message posted by: Trevor M. D'Souza