Patent ID: 8771955

Claim:
A method for quantifying and amplifying the amount of nucleic acids in a sample, comprising: (a) contacting nucleic acids from a sample with a nucleic acid amplification reaction mixture comprising a first fluorescent compound having a first fluorescence emission maximum, a second fluorescent compound having a second fluorescence emission maximum, and a third fluorescent compound having a third fluorescence emission maximum and a fluorescence intensity dependent on the concentration of amplified nucleic acids, wherein the first, second, and third fluorescence emission maxima are different, each having a fluorescence intensity dependent on the concentration of nucleic acids; (b) measuring the fluorescence of the first fluorescent compound to determine the amount of nucleic acids; (c) treating the nucleic acids and the nucleic acid amplification reaction mixture under conditions for amplifying the nucleic acids to provide amplified nucleic acids; and (d) measuring the fluorescence of the first fluorescent compound to determine the amount of amplified nucleic acids.