Massive parallel method for decoding DNA and RNA
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Abstract
This invention provides methods for attaching a nucleic acid to a solid surface and for sequencing nucleic acid by detecting the identity of each nucleotide analogue after the nucleotide analogue is incorporated into a growing strand of DNA in a polymerase reaction. The invention also provides nucleotide analogues which comprise unique labels attached to the nucleotide analogue through a cleavable linker, and a cleavable chemical group to cap the —OH group at the 3′-position of the deoxyribose.
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Citations
82 Claims
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1-60. -60. (canceled)
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61. A method of determining the identity of a nucleotide incorporated into a nucleic acid, comprising:
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a) providing a nucleic acid molecule attached to a solid surface, a polymerase, and four nucleotide analogues which are either (i) aA, aC, aG, and aT, or (ii) aA, aC, aG, and aU, wherein each nucleotide analogue is labeled with a unique label and contains a removable chemical moiety capping the 3′
-OH group;b) incorporating one of the nucleotide analogues into the nucleic acid with the polymerase; and c) detecting the unique label of the incorporated nucleotide analogue, so as to thereby determine the identity of the nucleotide incorporated into the nucleic acid. - View Dependent Claims (62, 63, 64, 65, 66, 67, 68, 69)
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70. A method of detecting the identity of nucleotides incorporated into nucleic acid, comprising:
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a) providing a plurality of nucleic acid molecules attached to a solid surface, a polymerase, and a plurality of nucleotide analogues, wherein each nucleotide analogue is labeled with a unique label and contains a chemical moiety capping the 3′
-OH group;b) incorporating a first nucleotide analogue into the nucleic acid with the polymerase; c) detecting the label of the incorporated nucleotide analogue; and d) removing the chemical moiety of the incorporated nucleotide analogue capping the 3′
-OH group, thereby permitting the further incorporation of the nucleotide analogues so as to create growing annealed strands of nucleic acid. - View Dependent Claims (71, 72, 73, 74, 75, 76)
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77. A plurality of nucleic acids immobilized on a solid surface, comprising incorporated nucleotide analogues, the nucleotide analogues each comprising a unique label and a chemical moiety capping the 3′
- -OH group.
- View Dependent Claims (78, 79, 80, 81, 82)
Specification