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Linked linear amplification of nucleic acids

  • US 6,027,923 A
  • Filed: 04/02/1997
  • Issued: 02/22/2000
  • Est. Priority Date: 07/23/1993
  • Status: Expired due to Fees
First Claim
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1. A process for amplifying a nucleic acid sequence of interest contained within a nucleic acid comprised of a first strand and a second strand that is complementary to the first strand, the process comprising:

  • (a) contacting each of the first and second strands with a primer that contains a non-replicable element and/or a cleavable element, under conditions such that first generation primer extension products are produced using said strands as templates, and wherein the primer for the first strand is selected such that a first generation primer extension product produced using the first strand as a template, when separated from the first strand, can serve as a template for synthesis of a second generation primer extension product of the primer for the second strand;

    (b) separating the first generation primer extension products from their respective templates to produce single-stranded molecules; and

    (c) treating the first generation primer extension products with the primers of step (a) under conditions such that second generation primer extension products are produced using the first generation primer extension products as templates;

    wherein the second generation primer extension products contain at least a portion of the sequence of the nucleic acid sequence of interest and no more than an insufficient portion of the binding site for the primer producing said first generation primer extension products.

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