Patent ID: 7208274
Filing Date: 2007-04-24
Classification: C12Q

Abstract:
1. A method for determining a sequence of alleles of a locus of interest, said method comprising: (a) amplifying 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 that cuts DNA at a distance from the recognition site and digestion with the restriction enzyme generates a 5′ overhang containing the locus of interest, and wherein the first primer contains a recognition site for a restriction enzyme that is different from the recognition site for the restriction enzyme on the second primer and contains a tag at the 5′ end; (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; (d) digesting the DNA of (c) with the restriction enzyme that recognizes the recognition site on the first primer; and (e) determining the sequence of the alleles of the locus of interest by determining the sequence of the digested DNA of (d) containing the labeled nucleotide.