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.
24 Citations
77 Claims
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1-60. -60. (canceled)
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61. A method of detecting a fluorescently labeled nucleotide incorporated into a DNA coupled to a phosphine moiety, comprising:
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a) reacting a DNA comprising an azido group with a compound comprising a phosphine moiety to produce the DNA coupled to the phosphine moiety; b) contacting the DNA coupled to the phosphine moiety of step a) with a polymerase and a fluorescently labeled nucleotide so as to incorporate the fluorescently labeled nucleotide into the DNA coupled to the phosphine moiety; and c) detecting the fluorescently labeled nucleotide incorporated into the DNA coupled to the phosphine moiety. - View Dependent Claims (62, 63, 64, 65, 66, 67)
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68. A method of detecting a fluorescently labeled nucleotide incorporated into a DNA fragment coupled to a phosphine moiety, comprising:
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a) reacting a DNA fragment having an azido group attached thereto through a linker with a compound comprising a phosphine moiety to produce the DNA fragment coupled to the phosphine moiety; b) contacting the DNA fragment coupled to the phosphine moiety of step a) with a polymerase and a fluorescently labeled nucleotide so as to thereby incorporate the fluorescently labeled nucleotide into the DNA fragment coupled to the phosphine moiety; and c) detecting the fluorescently labeled nucleotide incorporated into the DNA fragment coupled to the phosphine moiety. - View Dependent Claims (69, 70, 71, 72, 73, 74)
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75. A method of hybridizing, comprising:
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a) providing a nucleic acid template and a primer, the primer comprising an azido group; and b) hybridizing the primer to the nucleic acid template. - View Dependent Claims (76)
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77. A complex comprising a primer hybridized to a nucleic acid template, the primer comprising an azido group.
Specification