Patent ID: 8580508

Claim:
A method for real-time monitoring of nucleic acid amplification comprising: (a) amplifying a target nucleic acid, using a thermostable nucleic acid polymerase, 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; and ii) comprises a fluorophore; and said second probe; i) is capable of hybridizing to said first probe; ii) has a quencher molecule which quenches said first probe fluorophore when said first and second probes are hybridized to each other, wherein said fluorophore and said quencher are within fifteen base pairs of each other when said first probe and said second probe are hybridized to each other; and iii) is at least two nucleotides shorter in length than said first probe; and (b) detecting fluorescence of said first probe fluorophore in real-time to monitor amplification, wherein an increase in fluorescence correlates with amplification.