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Method and device for improved restriction fragment length polymorphism analysis

  • US 5,324,631 A
  • Filed: 08/26/1991
  • Issued: 06/28/1994
  • Est. Priority Date: 11/13/1987
  • Status: Expired due to Fees
First Claim
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1. A method for detecting variations in nucleic acid sequences, comprising(A) synthesizing a first primer which is complementary to a nonpolymorphic region of a genome of a test organism;

  • (B) synthesizing at least a second primer and a third primer, each of said second and third primers directly and detectably distinguishing between different sequence variations of a first polymorphic region of said genome, wherein said second and said third primers do not hybridize to the same DNA strand of said genome as said first primer;

    (C) labelling said second and third primers, respectively, with chromophores that are distinguishable, one from the other, in terms of fluorescent spectrum;

    (D) contacting all of said first, second and third primers with genomic DNA from said test organism under stringent hybridizing conditions, thereby to produce hybridization products;

    (E) amplifying said hybridization products, wherein said amplifying is accomplished by polymerase chain reaction to produce amplification products; and

    then(F) analyzing fluorescent spectra of said amplification products to determine which labels and by inference, which variations in said first polymorphic region of said genome are present.

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