Patent ID: 6642001
Filing Date: 2003-11-04
Classification: C12Q

Abstract:
A method of determining the identity of one or more nucleotides of interest in at least one nucleic acid sequence of interest comprising the steps of;a) combining i) a nucleic acid sequence of interest having at least one nucleotide of interest, ii) a detection probe comprising a detection sequence and a donor fluorescent molecule, iii) a primer comprising a variable nucleic acid sequence which hybridizes to said nucleic acid sequence of interest immediately adjacent to said nucleotide of interest, and a constant region comprising a nucleic acid sequence complementary to said detection sequence of the detection probe, and iv) one or more dideoxynucleotides labeled with an acceptor fluorescent molecule, under conditions suitable for template-dependent single base extension of said primer, wherein one of said dideoxynucleotides is added to said primer by template-directed extension, thereby forming a detection. complex comprising a primer extended by one labeled dideoxynucleotide and a detection probe, wherein the detection probe is hybridized to the constant region of said extended primer; and b) detecting the detection complex of a), thereby determining the identity of one or more nucleotides of interest in at least one nucleic acid sequence of interest.