Sequencing by Incorporation
First Claim
1. A method of sequencing a nucleic acid, comprising:
- (i) providing a first set of particles comprising at least one set of nucleic acid templates in a first microfluidic channel;
(ii) flowing a train of reagents comprising a plurality of sequencing reagents across the first set of particles, or flowing the first set of particles through the reagent train, thereby sequentially contacting the at least one set of nucleic acid templates with the plurality of sequencing reagents;
(iii) detecting a signal resulting from exposure of the first set of particles to the reagent train, thereby providing a portion of sequence of the nucleic acid template; and
,repeating steps (ii) and (iii), thereby sequencing the nucleic acid.
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Accused Products
Abstract
Nucleotides and nucleotide analogs are used in various sequencing by incorporation/sequencing by synthesis methods. Nucleotide analogs comprising 3′-blocking groups are used to provide reversible chain-termination for sequencing by synthesis. Typical blocking groups include phosphate groups and carbamate groups. Fluorescent nucleotides are used to perform sequencing by synthesis with detection by incorporation of the fluorescently labeled nucleotide, optionally followed by photobleaching and intercalating dyes are used to detect addition of a non-labeled nucleotide in sequencing by synthesis with detection by intercalation. Microfluidic devices, including particle arrays, are used in the sequencing methods.
87 Citations
25 Claims
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1. A method of sequencing a nucleic acid, comprising:
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(i) providing a first set of particles comprising at least one set of nucleic acid templates in a first microfluidic channel; (ii) flowing a train of reagents comprising a plurality of sequencing reagents across the first set of particles, or flowing the first set of particles through the reagent train, thereby sequentially contacting the at least one set of nucleic acid templates with the plurality of sequencing reagents; (iii) detecting a signal resulting from exposure of the first set of particles to the reagent train, thereby providing a portion of sequence of the nucleic acid template; and
,repeating steps (ii) and (iii), thereby sequencing the nucleic acid. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25)
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Specification