MiR-specific qPCR in different qPCR master mixes. A Comparison of amplification curves of a synthetic ssc-let-7d template in the ssc-let-7d miR-specific qPCR assay in QuantiFast and in Brilliant III qPCR Master mixes. B Melting curve analysis of the same experiment. No template control is labeled ntc. Melting curve analysis was performed from 60°C to 99°C. No change in fluorescence (dF/dT = 0) was observed above 80°C and this part of the curves was omitted from the figure. C Extrapolation of Cq as function of the log10 of the number of templates for the same experiment as in A was a straight line (R2 indicated on figure) and for both master mixes the PCR efficiency was 99% as calculated from the slope of the regression line.