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Rapid analysis of variations in a genome

  • US 20070196842A1
  • Filed: 12/11/2006
  • Published: 08/23/2007
  • Est. Priority Date: 03/01/2002
  • Status: Abandoned Application
First Claim
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1. A method for determining a sequence of alleles of a locus of interest, said method comprising:

  • (a) amplifying alleles of a locus of interest on a template DNA using a first and second primers, wherein the second primer contains a recognition site for a restriction enzyme such that digestion with the restriction enzyme generates a 5′

    overhang containing the locus of interest;

    (b) digesting the amplified DNA with the restriction enzyme that recognizes the recognition site on the second primer;

    (c) incorporating nucleotides into the digested DNA of (b), wherein;

    (i) a nucleotide that terminates elongation, and is complementary to the locus of interest of an allele, is incorporated into the 5′

    overhang of said allele, and (ii) a nucleotide complementary to the locus of interest of a different allele is incorporated into the 5′

    overhang of said different allele, and said terminating nucleotide, which is complementary to a nucleotide in the 5′

    overhang of said different allele, is incorporated into the 5′

    overhang of said different allele. (d) determining the sequence of the alleles of a locus of interest by determining the sequence of the DNA of (c).

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