Abstract
qBase, a free program for the management and automated analysis of qPCR data, is described
Highlights
Since its introduction more than 10 years ago [1], quantitative PCR has become the standard method for quantification of nucleic acid sequences
An important underlying assumption is that PCR efficiency is assay dependent and sample independent
With low M values (Additional data file 2: M ≅ 0.1), virtually identical results are obtained for the different selections of inter-run calibrators (IRCs) (Table 2)
Summary
Since its introduction more than 10 years ago [1], quantitative PCR (qPCR) has become the standard method for quantification of nucleic acid sequences. The software programs provided along with the various qPCR instruments allow for straightforward extraction of quantification cycle values from the recorded fluorescence measurements, and at best, interpolation of unknown quantities using a standard curve of serially diluted known quantities. These programs usually do not provide an adequate solution for the processing of these raw data (coming from one or multiple runs) into meaningful results, such as normalized and calibrated relative quantities. We developed an inter-run calibration algorithm to correct for (often underestimated) run-torun differences
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