Patent ID: 7632642

Claim:
A homogeneous detection method for at least one single-stranded amplification product of a non-symmetric nucleic acid amplification process that generates both double-stranded and single-stranded amplicons by extension of oligonucleotide primers by a DNA polymerase and that includes at least one primer-annealing temperature, comprising (a) forming an amplification reaction mixture that includes at least one nucleic acid target sequence and detection reagents, (b) amplifying said nucleic acid target sequence by said non-symmetric nucleic acid amplification process, (c) detecting double-stranded amplicons by a fluorescent signal, (d) detecting said at least one single-stranded amplification product by a sequence-specific fluorescent signal at a temperature below the at least one primer-annealing temperature, and (e) normalizing the sequence-specific fluorescent signal as a calculated ratio of fluorescence of said single-stranded amplification product to the fluorescence of said double-stranded amplicons.