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.
When there is a need to quickly analyze samples using a number of different PCR assays, it is likely that optimal conditions for each assay will not be the same. First, different assays often will require different annealing temperatures for their primers. In addition, amplicons may be designed to be of different lengths and therefore require varying durations of the extension step.READ MORE