Figure 1
Representative results for BRAF mutation screening using Sanger sequencing (A–D), single strand conformation analysis (SSCA; E–H), high resolution melting analysis (HRM; I–L) and competitive allele-specific TaqMan (CAST-PCR; M-P).
The first column is an example of wildtype BRAF, the second column of V600E, the third column of V600K and the fourth of K601E. Blue arrows indicate the wildtype profile, while red arrows indicate mutant profiles. For SSCA, E represents the V600E mutation and K the V600K mutation. In HRM graphs (difference plots), red lines indicate positive controls, blue lines negative controls, green lines the sample and black lines a sample/wildtype mix. In CAST PCR, green lines indicate the reference assay, blue lines the V600E assay and orange lines the V600K assay. Slight cross reactivity of V600E positive samples with the V600K assay (orange line) can be seen (N), but not the reverse (O). CAST-PCR using probes for V600E and V600K did not detect the K601E mutation (P).
