Patent ID: 6824980
Filing Date: 2004-11-30
Classification: C12Q

Abstract:
A method for detecting or quantifying a target nucleic acid in a sample comprising:(a) preparing at least one primer specifically matched to a predetermined position of the target nucleic acid; (b) annealing the at least one primer from (a) with the target nucleic acid to obtain a primer-nucleic acid duplex at the predetermined position of the target nucleic acid; (c) mixing the primer-nucleic acid duplex from (b) with a non-terminator nucleotide mixture consisting of: (i) dATP, dCTP, dGTP; (ii) dATP, dCTP, dTTP or dUTP; (iii) dATP, dGTP, dTTP or dUTP; or (iv) dCTP, dGTP, dTTP or dUTP wherein at least one non-terminator nucleotide of the non-terminator nucleotide mixture is labeled with a detectable marker;(d) performing isometric primer extension by enzymatic or chemical reaction in an appropriate buffer to form isometric primer extension products, wherein the primer extension terminates at a target nucleic acid nucleotide complementary to an omitted non-terminator nucleotide of (c); and (e) detecting or quantifying the amount of labeling signal on the isometric primer extension products.