Accurate and precise DNA quantification in the presence of different amplification efficiencies using an improved Cy0 method.

ABSTRACT: Quantitative real-time PCR represents a highly sensitive and powerful technology for the quantification of DNA. Although real-time PCR is well accepted as the gold standard in nucleic acid quantification, there is a largely unexplored area of experimental conditions that limit the application of the Ct method. As an alternative, our research team has recently proposed the Cy0 method, which can compensate for small amplification variations among the samples being compared. However, when there is a marked decrease in amplification efficiency, the Cy0 is impaired, hence determining reaction efficiency is essential to achieve a reliable quantification. The proposed improvement in Cy0 is based on the use of the kinetic parameters calculated in the curve inflection point to compensate for effici

SUBMITTER: Guescini M 

PROVIDER: S-EPMC3704541 | biostudies-literature | 2013

REPOSITORIES: biostudies-literature