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US 6,274,320 B1Granted
Method of sequencing a nucleic acid
AssigneeCuraGen Corporation
InventorsJonathan M. Rothberg, Joel S. Bader
Life of this patent
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Abstract
Disclosed herein are methods and apparatuses for sequencing a nucleic acid. The method includes annealing a population of circular nucleic acid molecules to a plurality of anchor primers linked to a solid support, and amplifying those members of the population of circular nucleic acid molecules which anneal to the target nucleic acid, and then sequencing the amplified molecules by detecting the presence of a sequence byproduct.
Claims32
A method for sequencing a nucleic acid, the method comprising:
- providing one or more or more nucleic acid anchor primers linked to a solid support, wherein said solid support includes at least one optical fiber;
- providing a plurality of single-stranded circular nucleic acid templates;
- annealing an effective amount of the nucleic acid anchor primer to at least one of the single-stranded circular templates to yield a primed anchor primer-circular template complex;
- combining the primed anchor primer-circular template complex with a polymerase to generate an extended anchor primer covalently linked to a nucleic acid comprising multiple copies of a sequence complementary to the circular nucleic acid template;
- annealing an effective amount of a sequencing primer to one or more copies of said covalently linked complementary nucleic acid;
- extending the sequencing primer with a polymerase and a predetermined nucleotide triphosphate to yield a sequencing product and, if the predetermined nucleotide triphosphate is incorporated onto the 3′ end of said sequencing primer, a sequencing reaction byproduct;
- and
- identifying the sequencing reaction byproduct, thereby determining the sequence of the nucleic acid.
5 more elements
Dependencies are inferred from the claim text.