LightCycler-based melting curve analysis. Depicted is the first derivation of the melting curve. The y-axis represents fluorescence intensity and the x-axis represents the temperature. Due to mismatches, mutations led to decreased melting temperatures of the hybridization probes from the amplification product. The peak at 52°C represents the wild-type allele, the peaks at 44°C represent mutated alleles. All mutated cases are heterozygous for the mutation and retain a wild-type allele, leading to double peaks in these cases.