Patent ID: 9689028
Date: 2017-06-27
CPC Classifications: C12Q

Claim:
1. A method for determining in a target sample the copy number of a first target nucleic acid and a second target nucleic acid, the method comprising: a) contacting a first target nucleic acid with a first primer set and a second target nucleic acid with a second primer set and adding a single detection label to form a reaction mixture in a homogenous system, wherein one of the primers of the first primer set is blocked from priming the first target nucleic acid and wherein one of the primers of the first primer set is blocked from priming the first target nucleic acid; wherein the primer blocked from priming the first target nucleic acid comprises a mismatched base at its 3′ end″ and wherein the first target nucleic acid comprises a tandem repent sequence; b) amplifying by polymerase chain reaction the first target nucleic acid with the first primer set to form a first amplicon having a first melting temperature (Tm) and amplifying by polymerase chain reaction the second target nucleic acid with the second primer set to form a second amplicon having a second Tm, wherein said second Tm is higher than said first Tm; c) during the polymerase chain reaction, determining the amount of the detection label at a first acquisition temperature, wherein said first signal acquisition temperature is below said first Tm; d) increasing the temperature of the reaction mixture to a second signal acquisition temperature and determining the amount of the detection label, wherein said second signal acquisition temperature is above said first TM and below said second Tm; e) repeating steps (b) through (d) at least one time; and f) determining the copy number of said first and said second target nucleic acids.