Patent ID: 8039216

Claim:
A method for identifying a nucleic acid in a sample, comprising: (a) providing a sample nucleic acid bound to a solid substrate; (b) contacting the sample nucleic acid with an oligonucleotide that is specific for the sample nucleic acid and is labeled with at least a first fluorescent dye; (c) contacting the sample nucleic acid with a second fluorescent dye that is different from the first fluorescent dye, wherein said second fluorescent dye interacts with the sample nucleic acid and is not covalently linked to an oligonucleotide; (d) amplifying the sample nucleic acid; (e) detecting the sample nucleic acid by observing fluorescence from the first fluorescent dye after said oligonucleotide hybridizes to said amplified sample nucleic acid; and determining the melting temperature of the amplified sample nucleic acid by measuring the fluorescence of the second fluorescent dye, and wherein there is no fluorescence resonance energy transfer (FRET) between the first fluorescent dye that labels the oligonucleotide and the second fluorescent dye.