Patent ID: 7312034
Filing Date: 2007-12-25
Classification: C12Q

Abstract:
1. A method of detecting each or any of a plurality of known, selected nucleotide target sequences, comprising: (a) mixing the target sequences with (i) a set of forward universal e-tag primers, each member of the set containing (ia) a target sequence that is complementary to one of the known selected target sequences, and (ib) an extension sequence which is unique to the target sequence of that member, (ii) one or more reverse primers that are complementary to said target sequences, and (iii) enzyme and nucleotide components of a primer extension reaction, to form a target-sequence reaction mixture; (b) reacting the reaction mixture under primer extension reaction conditions, to form extended target sequences, (c) reacting, under hybridization conditions, the extended target sequences with a set of electrophoretic tag (e-tag) probes, each member of the tag probe set having (i) an oligonucleotide portion that is complementary to one of said extension sequences, (ii) an electrophoretic probe having an electrophoretic mobility that is unique to a given extension sequence, and (iii) a linker joining the oligonucleotide portion and the electrophoretic probe, said linker being cleavable under selected conditions when the oligonucleotide portion of said probe is bound to a complementary target extension sequence, (d) treating the target sequences under said selected conditions, to release an e-tag reporter from each e-tag probe bound to a target sequence, (e) separating the released reporters electrophoretically, and (f) detecting separated released reporters, thereby to identify target sequences that hybridized to said probes.