Patent ID: 6376191
Filing Date: 2002-04-23
Classification: C12Q

Abstract:
A method for identifying sequence variations in a target polynucleotide in a sample as compared to a reference sequence, the method comprising the steps of:a) contacting an array of one or more groups of oligonucleotide primers with a reaction mixture containing the sample and reagents for polymerase-mediated polynucleotide amplification, wherein the array of oligonucleotide primers is immobilized on a solid phase support by the 5â€²-ends of the oligonucleotides, and further wherein each group of oligonucleotide primers is selected to span a particular region of the reference sequence, occupy a discrete area on the array, and comprise at least two sets of primers: 1) a first set that is exactly complementary to the reference sequence; and 2) one or more additional sets, each of which is identical to the first set of primers but for the nucleotide at the 3â€²-end which is different in each additional set; b) performing a plurality of cycles of polymerase-mediated polynucleotide amplification, whereby the target polynucleotide serves as template for the synthesis of detectable nascent polynucleotides which are extended from the sets of primers that are exactly complementary to the target polynucleotide, wherein all of the oligonucleotide primers of the array are present in the same amplification reaction; c) detecting the presence of synthesized polynucleotides which are captured on discrete areas of the solid phase support via corresponding immobilized primers; and d) identifying sequence variations in the at least one target polynucleotide according to a detected pattern of synthesized polynucleotides on the solid phase support.