Figure 5

Influence of amplification status on melting analysis. Panel A: The amplification plot of EGFR exon 19 shows insufficient amplification of several samples relative to the wild-type (blue). Panel B: The subsequent melting analysis of the same samples on a normalised plot shows the right shift of curves from the wild-type. Panel C: Dilution (1 in 20) of the wild-type (aqua) and the addition of 15 cycles allowed all samples to achieve similar levels of amplification. Panel D: The normalised plot now shows no aberrant right shifting of melting curves, identifying TX441 as mutation positive and TX454 and 07M708 as mutation negative.