Melanoma sample showing different results of the
p.V600E mutational analysis (case 30). Sanger sequencing as well as high resolution melting analysis show wildtype results (A and B, respectively). Pyrosequencing analysis resulted in a p.V600E mutation with a 5% mutant allele frequency having low relative fluorescent units of almost 30 (C). This result is confirmed by immunohistochemistry (D). Next generation sequencing resulted in a 2% mutant allele frequency with a coverage rate of 181 (E).