Patent ID: 7582429

Claim:
A method of PCR analysis comprising the steps of: mixing a dsDNA binding dye having a percent saturation of at least 90% with a sample comprising a selected target nucleic acid and primers configured for amplifying the selected target nucleic acid, amplifying the target nucleic acid in the presence of the dsDNA binding dye, and monitoring fluorescence of the dsDNA binding dye, wherein the monitoring step comprises melting the amplified target nucleic acid to generate a melting curve, repeating the mixing, amplifying, and generating a melting curve steps with at least one additional target nucleic acid, and comparing the melting curves, wherein the melting curve for the selected target nucleic acid is selected as the standard and is plotted as stranded across melting temperatures and the melting curve for each additional target nucleic acid is plotted as a difference from the stranded across the melting curves.