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Method for amplification and expression of nucleic acids in solid media and its application for nucleic acid cloning and diagnostics

DC CAFC
  • US 5,958,698 A
  • Filed: 08/17/1998
  • Issued: 09/28/1999
  • Est. Priority Date: 10/26/1992
  • Status: Expired due to Term
First Claim
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1. A method of detecting a nucleic acid sequence in a sample that may contain said sequence comprising the steps of:

  • (a) providing a cell-free, enzymatic, exponential amplification system;

    (b) forming a liquid mixture of the sample and said amplification system;

    (c) entrapping said liquid within solid surfaces comprising a thin layer;

    (d) incubating said trapped mixture under conditions promoting synthesis of an exponentially amplified nucleic acid product from said nucleic acid sequence; and

    (e) screening to detect said amplified product,wherein the average distance between the nearest solid surfaces is smaller than the distance which the synthesized nucleic acid product can migrate by diffusion during the reaction, andwherein copies of said nucleic acid sequence, if present in said sample, are sufficiently widely distributed in said liquid mixture to produce separate, detectable colonies of the synthesized nucleic acid product.

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