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Methods for controlling amplification

  • US 20090286286A1
  • Filed: 11/04/2008
  • Published: 11/19/2009
  • Est. Priority Date: 11/06/2007
  • Status: Abandoned Application
First Claim
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1. A method of amplifying nucleic acid on a solid support, comprising:

  • (a) providing i) a population of beads, each bead comprising one or more amplification primers, ii) a solution of amplification reagents comprising a thermostable polymerase, and iii) a population of nucleic acid template molecules,(b) mixing said beads and said template molecules in a first aliquot of said solution of amplification reagents so as to create a mixture;

    (c) treating the mixture under conditions such that at least a portion of said template molecules non-covalently bind to at least a portion of said beads to create bound template, and at least a portion of said primers on at least a portion of said beads are extended by said polymerase, so as to create treated beads;

    (d) manipulating said treated beads so as to remove at least a portion of said bound template so as to create manipulated beads; and

    (e) contacting said manipulated beads with a second aliquot of said solution of amplification reagents under conditions such that at least a portion of said extended primers is amplified to create loaded beads comprising immobilized amplified nucleic acid and unloaded beads lacking amplified nucleic acid.

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