Massive parallel method for decoding DNA and RNA
First Claim
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1. A method of sequencing nucleic acid, comprising:
- a) reacting a DNA strand containing an azido group with a phosphine;
b) growing a double-stranded DNA containing said DNA strand by incorporating a fluorescently labeled nucleotide into said strand; and
c) identifying the fluorescently labeled nucleotide, so as to sequence the nucleic acid.
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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.
227 Citations
20 Claims
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1. A method of sequencing nucleic acid, comprising:
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a) reacting a DNA strand containing an azido group with a phosphine; b) growing a double-stranded DNA containing said DNA strand by incorporating a fluorescently labeled nucleotide into said strand; and c) identifying the fluorescently labeled nucleotide, so as to sequence the nucleic acid. - View Dependent Claims (2, 3, 4, 5, 6, 7)
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8. A method of sequencing nucleic acid 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; 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; andd) identifying the fluorescently labeled nucleotide, so as to sequence the nucleic acid. - View Dependent Claims (9, 10, 11, 12, 13, 14)
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15. 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; b) growing a plurality of double-stranded DNA, each of which comprises one of said DNA strands, by incorporating a fluorescently labeled nucleotide; and c) identifying each fluorescently labeled nucleotide, so as to simultaneously sequence the plurality of different nucleic acids. - View Dependent Claims (16, 17, 18, 19, 20)
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Specification