DART-PCR (Data Analysis for Real-Time PCR) provides a simple means of analyzing real-time PCR data from raw fluorescence data (Peirson et al., 2003) (http://nar.oxfordjournals.org/cgi/content/full/31/14/e73/DCl). This allows an automatic calculation of amplification kinetics, as well as performing the subsequent calculations for the relative quantification and calculation of assay variability. Amplification efficiencies are also tested to detect anomalous samples within groups (outliers) and differences between experimental groups (amplification equivalence). Data handling was simplified by automating all calculations in an Excel® worksheet, and enables the rapid calculation of threshold cycles, amplification rate and resulting starting values, along with the associated error, from raw data. Differences in amplification efficiency are assessed using one-way analysis of variance (ANOVA), based upon the null hypotheses, that amplification rate is comparable within sample groups (outlier detection) and that amplification efficiency is comparable between sample groups (amplification equivalence) (Peirson et al, 2003).
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