Patent ID: 7824861

Claim:
A quantitative method for assaying the expression ratio between alleles differed by a single nucleotide polymorphism which comprises: (a) designing an allele-specific primer, according to nucleotide sequence of one allele, which has, in 3′ to 5′ order, a 3′-hydroxyl allele-specific nucleotide, 3′ perfect match nucleotides and 5′ perfect match nucleotides; wherein the allele-specific nucleotide is derived from known single nucleotide polymorphic site; (b) performing a polymerase chain reaction with an universal primer set which anneals to both alleles to obtain a total molecule number; (c) performing the polymerase chain reaction with an allele-specific primer and one of the universal primers to obtain the molecule number of one allele; and (d) calculating an expression frequency of one allele by molecular number of one allele/total molecular number.