Patent ID: 8093003

Claim:
A method for real-time monitoring of nucleic acid amplification comprising: (a) amplifying a target nucleic acid, using a thermostable nucleic acid polymerase having exonuclease activity, in the presence of a first oligonucleotide probe and a second oligonucleotide probe, said first probe; i) is capable of hybridizing to said target nucleic acid; ii) comprises a fluorophore; and iii) is not equal in length to said second probe; said second probe; i) is capable of hybridizing to said first probe; and ii) has a quencher molecule which quenches said first probe fluorophore when said first and second probes are hybridized to each other; and (b) detecting fluorescence of said first probe fluorophore in real-time to monitor amplification, wherein an increase in fluorescence correlates with amplification.