Massive parallel method for decoding DNA and RNA
AssigneeThe Trustees of Columbia University in the City of New York
InventorsJingyue Ju, Zengmin Li, John Robert Edwards, Yasuhiro Itagaki
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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.
Claims18
A nucleotide analogue which comprises:
- (a) a base which is an adenine, an analogue of adenine, a cytosine, an analogue of cytosine, a guanine, an analogue of guanine, a thymine, an analogue of thymine, a uracil, or an analogue of uracil;
- (b) a unique label attached through a cleavable linker to the base or to the analogue of the base;
- (c) a deoxyribose;
- (d) a —OR group at a 3′-position of the deoxyribose, wherein R is —CH2OCH3 or —CH2CH═CH2;
- and
- (e) a triphosphate group.
A nucleotide analogue which comprises:
- (a) a base which is an adenine, an analogue of adenine, a cytosine, an analogue of cytosine, a guanine, an analogue of guanine, a thymine, an analogue of thymine, a uracil, or analogue of uracil;
- (b) a unique label attached through a cleavable linker to the base or to the analogue of the base;
- (c) a deoxyribose;
- (d) a —OR group at a 3′-position of the deoxyribose, wherein R is a cleavable chemical group;
- and
- (e) a triphosphate group,
- wherein the nucleotide analogue has the structure:
- wherein Dye1, Dye2, Dye3, and Dye4 are four different dye labels.
Dependencies are inferred from the claim text.