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
80 Claims
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1-60. -60. (canceled)
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61. A method of sequencing nucleic acid, comprising:
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a) reacting a DNA strand containing an azido group with a phosphine; and b) growing a double-stranded DNA containing said DNA strand by incorporating a fluorescently labeled nucleotide into said strand. - View Dependent Claims (62, 63, 64, 65, 66, 67)
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68. A method comprising:
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a) providing a nucleic acid template hybridized to a primer, the primer comprising an azido group; b) reacting said azido group with a phosphine; and c) extending the primer hybridized to said nucleic acid template with a fluorescently labeled nucleotide or nucleotide analogue, wherein said fluorescently labeled nucleotide or nucleotide analogue has the label linked to the base and a blocking group on the 3′
-hydroxyl group.- View Dependent Claims (69, 70, 71, 72, 73, 74)
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75. A method of simultaneously sequencing a plurality of different nucleic acids, comprising:
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a) reacting each of a plurality of different DNA strands containing an azido group with a phosphine; and b) growing a plurality of double-stranded DNA, each of which comprises one of said DNA strands, by incorporating a fluorescently labeled nucleotide. - View Dependent Claims (76, 77, 78, 79, 80)
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Specification