Patent ID: 8088583

Claim:
A method of monitoring of a PCR amplification reaction, the method comprising: (a) preparing a PCR amplification reaction mixture by combining: (i) a template nucleic acid; (ii) forward and reverse nucleic acid primers; (iii) deoxyribonucleotides; (iv) a nucleic acid polymerase having 3′→5′ exonuclease activity; (v) a FET labeled probe, the FET labeled probe comprising a nucleic acid intercalator bonded to a FET labeled oligonucleotide, wherein the nucleic acid intercalator is covalently bonded to the 3′ end of the FET labeled oligonucleotide; and (b) subjecting the PCR amplification reaction mixture to PCR amplification conditions; (c) monitoring the reaction mixture for a fluorescent signal from the FET labeled oligonucleotide probe to obtain an assay result; and (d) employing the assay result to monitor the PCR amplification reaction.