Patent ID: 6500620
Filing Date: 2002-12-31
Classification: C12Q

Abstract:
A method for detecting multiple target polynucleotides in a sample, the method comprising the steps of:a) providing an array of oligonucleotide primers wherein each array contains at least two groups of oligonucleotide primers immobilized on discrete areas of a solid phase support, each group of oligonucleotide primers selectable for a particular target polynucleotide and comprising primer sequences complementary to a sequence of the particular target polynucleotide; b) contacting the array with a reaction mixture comprising a sample, said sample containing or suspected of containing multiple target polynucleotides, said reaction mixture further comprising reagents suitable for polynucleotide hybridization and amplification; c) performing a first round of polymerase-mediated polynucleotide amplification, whereby the target polynucleotides serve as initial templates for the synthesis of optionally detectable nascent polynucleotides which are extended from the immobilized primers; d) performing a second round of polymerase-mediated polynucleotide amplification in the presence of solution phase primers, wherein the solution phase primers serve as primers and the immobilized nascent polynucleotide strands from step c) serve as templates for the synthesis of detectable amplification products which are extended from the solution phase primers; and e) detecting the presence and quantity of the synthesized polynucleotides which are captured on discrete areas of the solid phase support via corresponding immobilized primers.