Use of reversible extension terminator in nucleic acid sequencing
First Claim
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1. A method for identifying a base in a template nucleic acid, comprising:
- (a) providing an optical confinement;
(b) providing a reaction mixture within the optical confinement, the reaction mixture comprising the template nucleic acid, a polymerase enzyme, and at least a first nucleotide analog, wherein the first nucleotide analog comprises a photolabile blocking group that terminates chain extension of a nascent nucleic acid strand when the first nucleotide analog is incorporated into the nascent nucleic acid strand; and
(c) identifying the base in the template nucleic acid strand by detecting the incorporated nucleotide analog.
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Abstract
The present invention relates to the uses of reversible extension terminator in conjunction with optical confinements in nucleic acid sequencing. The apparatus and methods embodied in the present invention are particularly useful for high-throughput and low-cost nucleic acid sequencing.
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Citations
51 Claims
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1. A method for identifying a base in a template nucleic acid, comprising:
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(a) providing an optical confinement; (b) providing a reaction mixture within the optical confinement, the reaction mixture comprising the template nucleic acid, a polymerase enzyme, and at least a first nucleotide analog, wherein the first nucleotide analog comprises a photolabile blocking group that terminates chain extension of a nascent nucleic acid strand when the first nucleotide analog is incorporated into the nascent nucleic acid strand; and (c) identifying the base in the template nucleic acid strand by detecting the incorporated nucleotide analog. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 29, 51)
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22. A system, comprising:
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an optical confinement; a reaction mixture disposed within the optical confinement, the reaction mixture comprising a template nucleic acid, a polymerase enzyme, and at least a first nucleotide analog, wherein the first nucleotide analog comprises a photolabile blocking group that terminates chain extension of a nascent nucleic acid strand when the first nucleotide analog is incorporated into the nascent nucleic acid strand. - View Dependent Claims (23, 24, 25, 26, 31)
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27. A method for identifying a base in a template nucleic acid, comprising:
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(a) providing a reaction mixture comprising the template nucleic acid, a polymerase enzyme, and at least a first nucleotide analog, wherein the first nucleotide analog comprises a removable blocking group that terminates chain extension of a nascent nucleic acid strand when the first nucleotide analog is incorporated into the nascent nucleic acid strand; (b) identifying the base in the template nucleic acid strand by detecting the incorporated nucleotide analog; (c) removing the removable blocking group from the incorporated nucleotide analog; and (d) repeating at least the identifying and removing steps with at least a second nucleotide analog without depleting at least one reagent from the reaction mixture of (a). - View Dependent Claims (28, 30, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50)
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Specification