Labelled nucleotides
DC CAFCFirst Claim
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1. A method of labeling a nucleic acid molecule, the method comprising incorporating into the nucleic acid molecule a nucleotide or nucleoside molecule, wherein the nucleotide or nucleoside molecule has a base that is linked to a detectable label via a cleavable linker and the nucleotide or nucleoside molecule has a ribose or deoxyribose sugar moiety, wherein the ribose or deoxyribose sugar moiety comprises a protecting group attached via the 2′
- or 3′
oxygen atom, and said protecting group can be modified or removed to expose a 3′
OH group and the protecting group comprises an azido group.
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Abstract
Nucleosides and nucleotides are disclosed that are linked to detectable labels via a cleavable linker group.
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Citations
15 Claims
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1. A method of labeling a nucleic acid molecule, the method comprising incorporating into the nucleic acid molecule a nucleotide or nucleoside molecule, wherein the nucleotide or nucleoside molecule has a base that is linked to a detectable label via a cleavable linker and the nucleotide or nucleoside molecule has a ribose or deoxyribose sugar moiety, wherein the ribose or deoxyribose sugar moiety comprises a protecting group attached via the 2′
- or 3′
oxygen atom, and said protecting group can be modified or removed to expose a 3′
OH group and the protecting group comprises an azido group. - View Dependent Claims (2, 3, 4, 5, 6, 8)
- or 3′
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7. A method of labeling a nucleic acid molecule, the method comprising incorporating into the nucleic acid molecule a nucleotide or nucleoside molecule, wherein the nucleotide or nucleoside molecule has a base that is linked to a detectable label via a cleavable linker and the nucleotide or nucleoside molecule has a ribose or deoxyribose sugar moiety, wherein the ribose or deoxyribose sugar moiety comprises a protecting group attached via the 2′
- or 3′
oxygen atom, and said protecting group can be modified or removed to expose a 3′
OH group and the protecting group and cleavable linker are removable under identical conditions. - View Dependent Claims (10, 11, 12, 13, 14)
- or 3′
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9. A method of labeling a nucleic acid molecule, the method comprising:
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a) incorporating into the nucleic acid molecule a nucleotide or nucleoside molecule, wherein the nucleotide or nucleoside molecule has a base that is linked to a detectable label via a cleavable linker, wherein the nucleotide or nucleoside molecule has a ribose or deoxyribose sugar moiety that comprises a protecting group attached via the 2′
or 3′
oxygen atom, and said protecting group can be modified or removed to expose a 3′
OH group and the protecting group comprises an azido group;b) detecting the detectable label and cleaving the cleavable linker, wherein the protected hydroxyl functionality and the protecting group are cleavable under identical conditions; and c) repeating steps a) and b) at least once.
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15. A method of labeling a nucleic acid molecule, the method comprising:
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a) incorporating into the nucleic acid molecule a nucleotide or nucleoside molecule, wherein the nucleotide or nucleoside molecule has a base that is linked to a detectable label via a cleavable linker, wherein the nucleotide or nucleoside molecule has a ribose or deoxyribose sugar moiety that comprises a protecting group attached via the 2′
or 3′
oxygen atom, and said protecting group can be modified or removed to expose a 3′
OH group and the protecting group comprises an allyl moiety;b) detecting the detectable label and cleaving the cleavable linker, wherein the protected hydroxyl functionality and the protecting group are cleavable under identical conditions; and c) repeating steps a) and b) at least once.
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Specification