Validation of the method for fluorescence-based quantitative real-time PCR (qPCR). Titration curves were obtained by using cDNA to create a series of dilutions, from 1× (no dilution) to 64 × dilution. Then the qPCR was run and log dilutions plotted against the obtained quantification cycle (Cq) values. Each Cq value was estimated as an average from 2 replicates and each point in the Figure represents mean ± SE from 3 separate cDNA samples. The linearity of the plots shows the equal amplification of the assay over a range of input DNA concentrations. These data were also used to estimate the efficiency (E) of the reaction for each gene of interest.