Chemically Modified Primers for PCR and Ligation Applications
Poster Jan 17, 2011
Elena Hidalgo Ashrafi, Sabrina Shore, Tony Le, Victor Timoshchuk, Natasha Paul, Richard Hogrefe, Inna Koukhareva, Alexandre Lebedev
PCR is an essential tool with utility in a variety of advanced applications. To improve the specificity of PCR, a unique approach to "Hot Start" PCR employing primers containing thermolabile modifications has been developed. These modified primers, named CleanAmp™ Primers, are amenable for use in Hot Start activation schemes as the modification is released after an initial denaturation step. CleanAmp™ Primers are available as either singly-modified CleanAmp™ Turbo or doubly-modified CleanAmp™ Precision. These two types of primers differ in the rate of release of unmodified primer thereby allowing for greater control of primer extension and an improvement in PCR amplification specificity. The faster deprotecting Turbo primers show a great advantage in multiplex PCR and low copy number detection. In reverse transcription PCR, the slower deprotecting Precision primers allow the user to perform reactions in a one-step, single tube format, reliably amplifying up to five targets simultaneously. The Precision primers also show benefit in the detection of ligation products by quantitative PCR, as they suppress nonspecific product formation for no template controls. Overall, this approach to "Hot Start" activation offers valuable improvements to PCR performance in multiple applications.
Despite the developments in conventional PCR, the complexity of multiplex Real Time PCR is still limited due to the lack of sufficient detection channels. To achieve high-end multiplexing capacity on standard Real Time PCR machines, Anapa Biotech has developed the MeltPlex® technology (see box on right).READ MORE