Patent ID: 6548247
Filing Date: 2003-04-15
Classification: C12Q

Abstract:
A method of detecting the presence of and determining the relative positions of at least two point mutations in a target polynucleotide, comprising:(a) hybridizing single-stranded oligonucleotide probes to a target polynucleotide to form hybrid, double-stranded polynucleotides such that mismatches occur at the sites of said point mutations, wherein said probes are complementary to a non-mutated sequence of said target polynucleotide and are labelled at one end but not both ends, and wherein said target polynucleotide is not labelled; (b) partially digesting the probe strand of said hybrid polynucleotide with a nucleic acid repair enzyme, wherein all possible recognition sites within the oligonucleotide probe are not cleaved and probe fragments of differing lengths are generated; (c) separating said probe fragments by size in a medium suitable for visualizing the separated probe fragments; and then (d) visualizing said separated probe fragments in said medium, whereby the presence and relative positions of said point mutations are determined.