Back to results
US 6,887,690 B2Granted
Dye-labeled ribonucleotide triphosphates
AssigneePE Corporation
InventorsPeter Virgil Fisher, Paolo Vatta, Shaheer H. Khan
Life of this patent
Estimating the term…
Abstract
The invention provides novel dye-labeled ribonucleotide analogs and methods for synthesizing those analogs. The compounds of the invention are especially useful for DNA sequencing by the polymerase chain reaction.
Claims40
A method for determining a polynucleotide sequence, comprising
- (i) annealing at least one primer to a template polynucleotide;
- (ii) extending said at least one primer in the presence of a mixture of at least four unlabeled dNTPs and at least one dye-labeled ribonucleotide having the formula:
- wherein B is a nucleobase;
- L is a linker;
- R3 is triphosphate, α-thiotriphosphate, or a salt thereof, and Dye is a reporter group;
- so that primer extension products that contain at least one dye-labeled ribonuoleotide are formed;
- (iii) cleaving one or more primer extension products to form a plurality of labeled fragments;
- (iv) separating the extension products by size;
- and
- (v) detecting the fragments to determine the polynucleotide sequence.
7 more elements
A method for detecting mutations in a polynucleotide, comprising
- annealing two primers to a template polynucleotide;
- extending the two primers in the presence of a mixture of at least four unlabeled dNTPs and at least one dye-labeled ribonucleotide having the formula:
- wherein B is a nucleobase;
- L is a linker;
- R3 is triphosphate, α-thiotriphosphate, or a salt thereof, and Dye is a reporter group;
- so that primer extension products that contain at least one dye-labeled ribonucleotide are formed;
- cleaving one or more primer extension products to form a plurality of labeled fragments;
- separating the fragments by size;
- and
- detecting the fragments to detect the mutations.
7 more elements
A method for determining a polynucleotide sequence, comprising
- (i) annealing at least one primer to a template polynucleotide;
- (ii) extending said at least one primer in the presence of a mixture of unlabeled dNTPs and at least one dye-labeled ribonucleotide having the formula:
- wherein B is a nucleobase;
- L is a linker;
- R3 is triphosphate, α-thiotriphosphate, or a salt thereof, and Dye is a reporter group;
- wherein at least one of the unlabeled dNTPs comprises a nucleobase that is the same as the nucleobase of at least one of the at least one dye-labeled ribonucleotide;
- so that primer extension products that contain at least one dye-labeled ribonucleotide are formed;
- (iii) cleaving one or more primer extension products to form a plurality of labeled fragments;
- (iv) separating the extension products by size;
- and
- (v) detecting the fragments to determine the polynucleotide sequence.
8 more elements
A method for detecting mutations in a polynucleotide, comprising
- annealing two primers to a template polynucleotide;
- extending the two primers in the presence of a mixture of unlabeled dNTPs and at least one dye-labeled ribonucleotide having the formula:
- wherein B is a nucleobase;
- L is a linker;
- R3 is triphosphate, α-thiotriphosphate, or a salt thereof, and Dye is a reporter group;
- wherein at least one of the unlabeled dNTPs comprises a nucleobase that is the same as the nucleobase of at least one of the at least one dye-labeled ribonucleotide;
- so that primer extension products that contain at least one dye-labeled ribonucleotide are formed;
- cleaving one or more primer extension products to form a plurality of labeled fragments;
- separating the fragments by size;
- and
- detecting the fragments to detect the mutations.
8 more elements
Dependencies are inferred from the claim text.