Principle of high-resolution melting curve analysis (HRM) for detection of allelic expression imbalance. A single labelled fluorescent probe is designed with complete complementarity to one allele of the exonic SNP chosen as marker, while mismatching the other allele. Following an asymmetric PCR reaction in presence of the probe, HRM analysis allows the alleles in heterozygous individuals to be distinguished by differences in their melting temperatures (Tm), with a fluorescent signal correlated to the relative abundance of each transcript. The Allele 2/Allele 1 ratio is calculated as h2/h1.
Nguyen-Dumont et al. BMC Medical Genomics 2011 4:39 doi:10.1186/1755-8794-4-39