Patent ID: 6300070
Filing Date: 2001-10-09
Classification: C12Q

Abstract:
A solid-phase, multi-stage method of amplifying one, or more, target nucleic acid molecules comprising two or more stages of bridge amplification, wherein single-stranded amplification molecules produced in the first stage of bridge amplification initiate a second stage of bridge amplification, and each subsequent stage of bridge amplification is initiated with single-stranded amplification molecules produced in the previous stage of bridge amplification, wherein each stage of bridge amplification comprises the steps of:(a) hybridizing target nucleic acid molecules to a set of oligonucleotide primers immobilized on a solid support;(b) thermocycling to amplify the target nucleic acid molecules hybridized to the oligonucleotide primers by the formation of bridge amplification double-stranded molecules;(c) cleaving and denaturing the bridge amplification double-stranded molecules to form single-stranded amplification molecules;(d) recovering the single-stranded amplification molecules of step (c);(e) contacting the single-stranded amplification molecules with a fresh solid support; and(f) repeating steps (a) through (e) to amplify the target nucleic acid molecules.