Figure 3. Calculation of the optimal number of reference genes for normalization by geNormPlus.

geNormPlus calculates the optimal number of reference genes taking into account the variable V as the average pairwise variation between two sequential candidate reference miRNAs. The dotted line illustrates the cut-off value 0.15. In our cohort the optimal number of reference genes was two (V2/3) for Pool A (white boxes) and four (V4/5) for Pool B (grey boxes indicate a V-value above the cut-off).