Fig 3. Dynamic range of BRAF V600E SNPase-ARMS qPCR.
Exemplary qPCR amplification plots of a serial dilution of 105 to ten BRAF V600E copies in a background of 2 × 105 (A) and 105 (B) wild type BRAF copies (following a 15 cycle SNPase preamplification step) are shown. The respective target-copy number is indicated in the plot. Delta R (y-axis) is plotted against quantification cycle (x-axis). qPCR threshold level is represented by the grey horizontal line. All reactions containing target DNA (blue) are positive and quantifiable with a single negative at ten copies in (A). Negative control samples (orange) show delayed amplification of approximately seven quantification cycles or do not amplify at all. No signal amplification was observed in the NTC sample wells. Results of wells containing three or one target copy are shown in Fig 4 and S3 Fig.