Figure 3

The influence of amplicon choice on mutation detection. The top panels show difference curves and melting peak curves for the original BRCA1 exon 7 amplicon which did not readily detect single base insertions or deletions. While the c.427G>T (red) and the c.314A>G (green) mutations are readily detectable in both visualisations, the pathogenic c.329insA (navy) and c.302-2delA (yellow) single base pair insertion and deletion mutations were difficult to detect as they melt like the wildtype controls (grey). The original amplicon was then divided into two overlapping amplicons. The bottom panels show difference curves and melting peak curves for BRCA1 amplicon exon 7A which detects the single base insertion and deletion. The c.329insA (navy) and c.302-2delA (yellow) clearly differ from the wildtype controls (grey) in the shorter amplicon. The BRCA1 c.314A>G (dark green) mutation is also readily detectable in both visualisations.