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Plasmid-based mutation detection system in transgenic fish

  • US 6,472,583 B1
  • Filed: 10/26/1999
  • Issued: 10/29/2002
  • Est. Priority Date: 10/26/1998
  • Status: Expired due to Term
First Claim
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1. A method for detecting mutations in the DNA of a transenic fish comprising:

  • providing a transgenic fish, wherein the DNA of the transgenic fish comprises multiple concatemeric genomically integrated copies of a plasmid comprising an assayable mutation target nucleic acid sequence;

    gently disaggregating at least a portion of the fish to yield disaggregated fish material;

    digesting the disaggregated fish material with a proteinase for a period of no longer than about 1-½

    hours at a temperature of about 37°

    C.;

    extracting DNA comprising the mutation target nucleic acid sequence from the disaggregated fish material sufficient to detect a mutation in the mutation target nucleic acid sequence; and

    detecting the presence of a mutation in the mutation target nucleic acid sequence.

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