FIELD: biochemistry.
SUBSTANCE: methods involves contacting a target nucleic acid molecule under substantially isothermal conditions with a polymerase, two or more primer/template 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/template oligonucleotides comprises one or more oligonucleotide primers or several 2′-modified nucleotides, in the 3′-end of sequence, complementary to the target nucleic acid molecule; generating amplicons comprising at least a portion of said target nucleic acid molecule; 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.
EFFECT: described are methods for quantitative determination of specific product in an amplification reaction with nicking and extension and control method monitoring in real time a nicking and extension amplification reaction.
13 cl, 10 dwg, 3 ex
