Patent ID: 7829312

Claim:
A method of assessing an amount of a known target nucleic acid sequence in a sample comprising: a) co-amplifying using a duplex PCR said known target nucleic acid sequence and a known amount of a known control nucleic acid sequence using different target and control specific primers to produce respective target and control amplicons, wherein said control nucleic acid sequence is different than said target nucleic acid sequence, and wherein said co-amplifying is stopped during an exponential phase of the duplex PCR; b) predetermining a dispensation order of nucleotides based on the nucleotide sequence order of the target nucleic acid and control nucleic acid sequence; c) dispensing the nucleotides in a single reaction in the predetermined dispensation order in a primer extension reaction such that the target and control nucleic acids are extended sequentially; and d) determining relative amounts of said target and control amplicons by comparing a quantity of nucleotides incorporated during said primer extension reaction for said target amplicon with a quantity of nucleotides incorporated during said primer extension reaction for said control amplicon, wherein relative amounts of said respective amplicons are proportional to relative quantities of nucleotides incorporated during said primer extension reactions and said amount of said target nucleic acid sequence in said sample is proportional thereto.