Patent ID: 9587267
Date: 2017-03-07
CPC Classifications: C12Q

Claim:
1. A method for quantifying nucleic acid in a sample, comprising: (a) providing a sample comprising nucleic acid to be quantified; (b) adding a sufficient amount of a single-stranded oligonucleotide probe having a length of about 70 to about 100 nucleotides to the sample, thereby forming a probe-nucleic acid sample, the single-stranded oligonucleotide probe comprising a hybridization sequence and a 5′ tag sequence, wherein the hybridization sequence can specifically hybridize to the nucleic acid to be quantified with at least 90% complementarity, and wherein the 5′ tag sequence is located 5′ of the hybridization sequence; (c) incubating the probe-nucleic acid sample from step (b) under conditions which allow hybridization of the hybridization sequence of the single-stranded oligonucleotide probe to the nucleic acid to be quantified, thereby forming a hybridized nucleic acid sample; (d) incubating the hybridized nucleic acid sample from step (c) under conditions which allow extension of the single-stranded oligonucleotide probe hybridized to the nucleic acid to be quantified, thereby forming an extended oligonucleotide probe; (e) after step (d), removing any of the single-stranded oligonucleotide probe added in step (b) and not hybridized to the nucleic acid to be quantified in step (c); and (f) following step (e), measuring an amount of the nucleic acid to be quantified in the sample by quantifying the extended oligonucleotide probe, wherein said quantifying the extended oligonucleotide probe comprises amplifying the 5′ tag sequence of the extended oligonucleotide probe using a primer that can hybridize to the 5′ tag sequence.