Patent ID: 8114595

Claim:
A method of detecting a target nucleic acid in a sample, the method comprising: (a) providing at least one labeled oligonucleotide, which oligonucleotide is labeled with at least one light emitting moiety and wherein at least a subsequence of the labeled oligonucleotide is sufficiently complementary to at least a subsequence of at least one target nucleic acid and/or to at least a subsequence of an amplicon of the target nucleic acid such that the labeled oligonucleotide hybridizes with the target nucleic acid and/or the amplicon of the target nucleic acid under at least one selected condition; (b) providing at least one soluble light emission modifier, selected from a diazine dye or a thiazine dye, in solution in free form that non-covalently associates with the labeled oligonucleotide and that modifies a light emission from the labeled oligonucleotide to a greater extent than from a labeled fragment of the oligonucleotide at a temperature of at least 40° C.; (c) amplifying the nucleic acid in the sample in the presence of the labeled oligonucleotide and the soluble light emission modifier in an amplification reaction that comprises the selected condition such that the labeled oligonucleotide, hybridized with the target nucleic acid or the amplicon of the target nucleic acid, is cleaved to produce at least one labeled oligonucleotide fragment; and, (d) detecting light emission at least from the labeled oligonucleotide fragment during (c), thereby detecting the target nucleic acid.