Abstract: A method of determining whether one or more forms of a nucleic acid analyte are present in a sample. The method includes dissolving an amplification reagent with a first solvent, where the amplification reagent contains oligonucleotides sufficient to amplify and detect a first region of a first form of the analyte, where the first solvent contains one or more oligonucleotides which, in combination with the oligonucleotides of the amplification reagent, are sufficient to amplify and detect a second region of a second form of the analyte, where the one or more oligonucleotides of the first solvent are insufficient to amplify and detect the first or second form of the analyte, and where the first and second regions each comprise a different nucleotide base sequence.

Abstract: The present disclosure relates to compositions and methods, and related systems, products and kits, for performing temperature-dependent multiplex invasive cleavage assays, where a plurality of target nucleic acids are detected and distinguished from each other in a procedure using only a single fluorescent moiety as the reporter, and single channel fluorescence detection. In some embodiments, at least one of the plurality of target nucleic acids is an amplified nucleic acid, where progress in a thermal cycling amplification reaction is monitored as a function of time (i.e., real-time amplification).