DESIGN OF STEM-LOOP PROBES AND UTILIZATION IN SNP GENOTYPING
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United States of America Patent
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Issued Date -
Apr 18, 2013
app pub date -
Oct 13, 2011
filing date -
Oct 13, 2011
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Abstract
Stem-loop probe for single nucleotide polymorphism (SNP) genotyping of individual SNP nucleic acid target sequences has first, second, and third single stranded nucleic acid portions. Second portion is between the first and third portions. First and third portions build a double stranded, intramolecular stem. Second portion forms a single stranded oligonucleotide loop with a nucleotide sequence, complementary to individual SNP nucleic acid target sequences. The nucleotide sequence of probe is matches probe/target hybrids have a melting point Tm that is at least 5° C. higher than Tm of mismatched probe/target hybrids. A method of detecting SNP in nucleic acid containing samples uses a pair of stem-loop probes for SNP genotyping of two individual SNP nucleic acid target sequences. The probes comprise the same first, second, and third portions and a ratio of perfect match probe/target hybrids to mismatched probe/target hybrids is detected at a certain temperature.
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