Figure 4

Multiplexing 3 genetic markers in one MT-PCR-HRM screening reaction. First order differential plots of a High Resolution Melt derived from a multiplex MT-PCR-HRM experiment (three replicates per treatment, green lines) performed on BAC matrix pools (5 × 384 well plates) using three independent loci as revealed on a Qiagen Rotor-Gene 6000. Positive control samples were generated using individual BAC clones containing each of the three loci (M106, red line; M34, orange line and M45, purple line). Negative control (blue line) lacked template DNA. Note that the positive control traces each occupy a diagnostic position on the trace and that the three experimental traces yield peaks corresponding to each of these positions (green).