Analysis of SNP rs2104772 by High-Resolution Melt analysis (HRM).

A-C) Line graph showing the detected or derived fluorescence intensity for the analyzed SNP (i.e. melting curves). The displayed examples included measurements for A/A homozygotes (red line, n = 5)) and T/T homozygotes (blue line, n = 3) relative to the heterozygote A/T (green line, n = 12). Every sample was analyzed in duplicate. A) Raw data of the pre-melt, melt, and post-melt regions. B) Normalized data derived from the raw data plots. C) Melting curves derived after normalization versus the A/A genotype. D-F) Sequence analysis of the three identified genotypes in chromatograms presenting the forward sequence. Arrows link the single-nucleotide polymorphism (SNP) position 44513. The presence of 'W' in A/T genotype denotes heterozygosis for the SNP where a double-peak is present at position 44513 for both sequenced alleles (arrow, nucleotides A and T simultaneously).