Comparison of Analytic Methods for Quantitative Real-Time Polymerase Chain Reaction Data

摘要

Abstract Polymerase chain reaction (PCR) is a laboratory procedure to amplify and simultaneously quantify targeted DNA molecules, and then detect the product of the reaction at the end of all the amplification cycles. A more modern technique, real-time PCR, also known as quantitative PCR (qPCR), detects the product after each cycle of the progressing reaction by applying a specific fluorescence technique. The quantitative methods currently used to analyze qPCR data result in varying levels of estimation quality. This study compares the accuracy and precision of the estimation achieved by eight different models when applied to the same qPCR dataset. Also, the study evaluates a newly introduced data preprocessing approach, the taking-the-difference approach, and compares it to the currently used approach of subtracting the background fluorescence. The taking-the-difference method subtracts the fluorescence in the former cycle from that in the latter cycle to avoid estimating the background fluorescence. The..." /> rel="meta" type="application/atom+xml" href="http://dx.doi.org/10.1089%2Fcmb.2015.0023" /> rel="meta" type="application/rdf+json" href="http://dx.doi.org/10.1089%2Fcmb.2015.0023" /> rel="meta" type="application/unixref+xml" href="http://dx.doi.org/10.1089%2Fcmb.2015.0023" /> 展开▼