Web-based LinRegPCR: application for the visualization and analysis of (RT)-qPCR amplification and melting data.

ABSTRACT:

Background

The analyses of amplification and melting curves have been shown to provide valuable information on the quality of the individual reactions in quantitative PCR (qPCR) experiments and to result in more reliable and reproducible quantitative results.

Implementation

The main steps in the amplification curve analysis are (1) a unique baseline subtraction, not using the ground phase cycles, (2) PCR efficiency determination from the exponential phase of the individual reactions, (3) setting a common quantification threshold and (4) calculation of the efficiency-corrected target quantity with the common threshold, efficiency per assay and C_q per reaction. The melting curve analysis encompasses smoothing of the observed fluorescence data, normalization to remov

SUBMITTER: Untergasser A 

PROVIDER: S-EPMC8386043 | biostudies-literature | 2021 Aug

REPOSITORIES: biostudies-literature