REAL TIME PCR DETECTION OF SINGLE NUCLEOTIDE POLYMORPHISMS
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United States of America Patent
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Issued Date -
Apr 12, 2012
app pub date -
Jun 13, 2011
filing date -
Oct 7, 2010
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Abstract
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.
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