Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Preparing compound containing saccharide radical
Patent
1999-09-18
2000-08-22
Sisson, Bradley L.
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Preparing compound containing saccharide radical
435 6, 435183, 435810, 436172, 536 221, 536 243, 536 2433, 536 253, C12P 1934
Patent
active
061070610
ABSTRACT:
The present invention relates to assays for the detection of nucleotide bases at predetermined locations on polynucleotides of interest. The embodiments of the invention include techniques relating to methods in which a primer extension reaction is designed to only extend a single nucleotide base. The invention, in part, relates to the improvement of adding corresponding non-labeled chain terminators to a primer extension reaction so as to increase the accuracy and reliability of nucleotide base determinations made by the assay. The invention is particularly useful when applied to assay methods in which the primer extension reaction is detected by a fluorescent dyes conjugated to chain terminators incorporated in the extension reactions. A first embodiment of the invention relates to methods for identifying a nucleotide base at the predetermined location on a polynucleotide of interest. Other embodiments of the invention include compositions for identifying a nucleotide base at a predetermined location on a polynucleotide of interest. One embodiment of such a composition is a mixture comprising a polynucleotide for analysis, an extendable polynucleotide that can hybridize of a specific location on the polynucleotide for analysis, a first labeled terminator, a second labeled terminator, and an unlabeled terminator corresponding of the first labeled terminator. The compositions may comprise additional labeled terminators and corresponding unlabeled terminators. Another embodiment of the subject compositions is a mixture comprising at least two labeled terminators and one or more corresponding unlabeled terminators. The compositions may comprise additional labeled terminators and corresponding unlabeled terminators. Other embodiments of the invention include kits for identifying a nucleotide base at a predetermined location on the polynucleotide of interest.
REFERENCES:
patent: 5625081 (1997-04-01), Trainor
patent: 5925520 (1999-07-01), Tully et al.
Bortner Scott R.
Sisson Bradley L.
The Perkin-Elmer Corporation
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