Methods of producing and sequencing modified polynucleotides
First Claim
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1. A method for producing a modified polynucleotide sequence comprising:
- a) annealing at least one primer to a template polynucleotide sequence;
b) extending said primer in the presence one or more nucleoside triphosphates,wherein at least one of the nucleoside triphosphates is a modified nucleoside triphosphate comprising a general structure [I];
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Abstract
The present invention encompasses methods for producing a modified polynucleotide sequence that comprises a (e.g., one or more) phosphorothiolate linkage, methods for determining a polynucleotide sequence comprising a (e.g., one or more) phosphorothiolate linkage, and methods for separating forward and reverse extension products that comprise a (e.g., one or more) phosphorothiolate linkage. The invention also encompasses kits for producing and/or determining the sequence of a modified polynucleotide that comprises a (e.g., one or more) phosphorothiolate linkage.
11 Citations
19 Claims
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1. A method for producing a modified polynucleotide sequence comprising:
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a) annealing at least one primer to a template polynucleotide sequence; b) extending said primer in the presence one or more nucleoside triphosphates, wherein at least one of the nucleoside triphosphates is a modified nucleoside triphosphate comprising a general structure [I]; - View Dependent Claims (2, 3, 4, 5, 6)
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7. A method for producing a modified polynucleotide sequence comprising:
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a) annealing at least one primer to a template polynucleotide sequence; b) extending said primer in the presence of a mixture of nucleoside triphosphates, wherein at least one of the nucleoside triphosphates is a modified nucleoside triphosphate comprising a general structure [III]; - View Dependent Claims (8, 9, 10, 11)
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12. A method for determining a polynucleotide sequence comprising:
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a) annealing a plurality of primers to a plurality of template polynucleotide sequences; b) extending the plurality of primers in the presence of one or more nucleoside triphosphates, wherein at least one of the nucleoside triphosphates is a modified nucleoside triphosphate comprising a general structure [I]; - View Dependent Claims (13, 14, 15, 16)
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17. A method for determining a polynucleotide sequence comprising:
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a) annealing a plurality of primers to a plurality of template polynucleotide sequences, wherein each primer in the plurality of primers comprises a first tag; b) extending the plurality of primers in the presence of one or more nucleoside triphosphates, wherein at least one of the nucleoside triphosphates is a modified nucleoside triphosphate comprising a general structure [III]; - View Dependent Claims (18, 19)
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Specification