Quantification of siRNA by a Novel Competitive-qPCR Method
Poster Sep 29, 2007
We have developed a competitive qPCR method in which siRNA competes with a homologous forward primer to bind template DNA, giving siRNA concentration dependent inhibition. The addition of E6-siRNA to cqPCR led to inhibition of amplification in a linear concentration-dependent manner, with as little as 200pg of siRNA capable of being detected. Irrelevant siRNA had no effect on amplification confirming specificity.
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