A novel data processing method CyC* for quantitative real time polymerase chain reaction minimizes cumulative error.

ABSTRACT: Quantitative real-time polymerase chain reaction (qPCR) is routinely conducted for DNA quantitative analysis using the cycle-threshold (Ct) method, which assumes uniform/optimum template amplification. In practice, amplification efficiencies vary from cycle to cycle in a PCR reaction, and often decline as the amplification proceeds, which results in substantial errors in measurement. This study reveals the cumulative error for quantification of initial template amounts, due to the difference between the assumed perfect amplification efficiency and actual one in each amplification cycle. The novel CyC* method involves determination of both the earliest amplification cycle detectable above background ("outlier" C*) and the amplification efficiency over the cycle range from C* to the next two

SUBMITTER: Zhang L 

PROVIDER: S-EPMC6559663 | biostudies-literature | 2019

REPOSITORIES: biostudies-literature