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Fluorescent in situ rt-pcr

  • US 20030059801A1
  • Filed: 09/18/2002
  • Published: 03/27/2003
  • Est. Priority Date: 12/09/1999
  • Status: Active Grant
First Claim
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1. A method for in situ amplification of a target nucleic acid in a selected cell comprising:

  • a) contacting a fixed, permeabilized cell with a restriction endonuclease composition comprising at least one restriction endonuclease to produce a restriction digest;

    b) contacting said cell with a DNase to produce a DNase digested cell;

    c) incubating said cell with a reverse transcriptase (RT) cocktail comprising an RT enzyme and a RT primer specific for said target nucleic acid to produce a cDNA; and

    d) amplifying said cDNA using a PCR reaction in the presence of forward and reverse PCR primers specific for said target nucleic acid wherein at least one of said PCR primers is labeled.

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