Patent ID: 7445898

Claim:
A method of quantifying a target nucleic acid in an amplification reaction, the method comprising incubating an amplification reaction that comprises a template comprising the target nucleic acid with: amplification primers, a polymerase, an enzyme having 3′ to 5′ exonuclease activity, and a probe in which the 3′ end of the probe has the sequence TCAGCN, TCAGCTN, or TCANNN, wherein each of the N residues represents a mismatch to the target nucleic acid sequence, where the amplification reaction is incubated under conditions in which i) the amplification primers are extended by the polymerase to amplify the target nucleic acid; ii) the probe specifically hybridizes to the target nucleic acid, wherein the 3′ nucleotide is cleaved from the probe; and detecting the amount of cleavage product, thereby quantifying the target nucleic acid.