Patent ID: 9096897
Filing Date: 2015-08-04
Classification: C12Q

Abstract:
1. A method of quantifying a specific product in a nicking and extension amplification reaction, the method comprising: (a) contacting a target nucleic acid molecule under substantially isothermal conditions with a polymerase, two or more primer oligonucleotides, each of which specifically binds to a complementary sequence on the target nucleic acid molecule, a nicking enzyme, and a detectable polynucleotide probe, wherein each of the primer oligonucleotides comprises one or more 2′ modified nucleotides positioned at the 3′ end of the sequence complementary to the target nucleic acid molecule; (b) generating amplicons comprising at least a portion of said target nucleic acid molecule; and (c) detecting a signal specific for oligonucleotide probe hybridization to the target nucleic acid molecule or amplicon thereof, wherein the signal indicates the quantity of the target nucleic acid molecule present in the sample or an amplicon thereof.