REAL TIME PCR DETECTION OF SINGLE NUCLEOTIDE POLYMORPHISMS

摘要

Disclosed are methods and kits for the detection of a polymorphism during real-time PCR. Real-time PCR amplification of a target nucleic acid sequence is performed using PCR primer primers that anneal to sequences flanking a single nucleotide polymorphism (SNP) of interest. The real-time PCR reaction includes a labeled probe comprising a RNA sequence that is designed to anneal to DNA sequences at the location of the SNP. An RNA:DNA heteroduplex can then form between the SNP in the PCR fragment and the probe's RNA sequences that are complementary to the SNP. RNase H cleavage of the RNA sequence in the RNA:DNA heteroduplex results in increase in intensity of the signal generated from the label that is indicative of the presence of an SNP in the target nucleic acid.