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Nucleic acid amplification methods

  • US 6,593,086 B2
  • Filed: 12/01/2000
  • Issued: 07/15/2003
  • Est. Priority Date: 05/20/1996
  • Status: Expired due to Term
First Claim
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1. A method for detection of a target nucleic acid within a cell comprising the steps of:

  • (a) embedding the cell to be analyzed for the presence of the target nucleic acid within a matrix;

    (b) adding a circularizable Amp-probe-2 having 3′ and

    5′

    regions that are complementary to adjacent but noncontiguous sequences in the target nucleic acid, said 3′ and

    5′

    regions separated by a linker region that is neither complementary nor hybridizable to a nucleotide sequence in the target nucleic acid such that a complex is formed comprising the target nucleic acid and Amp-probe-2;

    (c) ligating the 3′ and

    5′

    ends of said circularizable Amp-probe-2 with a ligating agent that joins nucleotide sequences such that a circular Amp-probe-2 is formed;

    (d) washing said complex; and

    (e) detecting said complex, wherein the detection thereof indicates the presence of the target nucleic acid in the embedded cell.

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