Quantification of siRNA by a Novel Competitive-qPCR Method
Abstract
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.
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.
