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High-throughput screening method for identification of genetic mutations or disease-causing microorganisms using segmented primers

  • US 5,888,778 A
  • Filed: 06/16/1997
  • Issued: 03/30/1999
  • Est. Priority Date: 06/16/1997
  • Status: Expired due to Term
First Claim
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1. A method for screening genomic nucleic acid samples to identify a nucleic acid mutation comprising the steps of:

  • (i) isolating a plurality of genomic nucleic acids;

    (ii) exposing said nucleic acids to one or more segmented primers, each comprising a first primer having a length between 5 and 8 nucleotides, and a second primer having a length between 15 and 100 nucleotides and a 3'"'"' non-extendible nucleic acid, wherein said first and second primers hybridize to regions of a target nucleic acid that are separated by no more than one nucleotide on said target and wherein said second primer binds upstream of said first primer;

    (iii) conducting a template-based nucleic acid extension reaction, thereby to add a single terminal nucleotide to one or more of said first probes;

    (iv) isolating extended first probes; and

    (v) determining the sequence of said extended first probes;

    thereby to detect the presence or absence of a genetic alteration.

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