Labelled nucleotides
First Claim
1. A method for determining the identity of a nucleotide in a target single-stranded polynucleotide, comprising:
- (a) providing one or more of the nucleotides A, G, C and T or U in which each of said nucleotides has a base that is attached to a distinct detectable label via a linker, said linker being cleavable with a water-soluble phosphine and said cleavable linker comprising a disulfide; and
a nascent polynucleotide complementary to the target polynucleotide, one of said provided nucleotides being suitable for incorporation into said nascent polynucleotide;
(b) incorporating the nucleotide suitable for incorporation into said nascent polynucleotide;
(c) detecting the detectable label of the incorporated nucleotide to identify the incorporated nucleotide; and
(d) cleaving the disulfide linker of the incorporated nucleotide with a water-soluble phosphine.
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Accused Products
Abstract
The invention provides a nucleotide or nucleoside having a base attached to a detectable label via a cleavable linker, characterized in that the cleavable linker contains a moiety selected from the group comprising: Formula (I) (wherein X is selected from the group comprising O, S, NH, and NQ wherein Q is a C1-10 substituted or unsubstituted alkyl group, Y is selected from the group comprising O, S, NH, and N(allyl), T is hydrogen or a C1-10 substituted or unsubstituted alkyl group and * indicates where the moiety is connected to the remainder of the nucleotide or nucleoside).
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Citations
13 Claims
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1. A method for determining the identity of a nucleotide in a target single-stranded polynucleotide, comprising:
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(a) providing one or more of the nucleotides A, G, C and T or U in which each of said nucleotides has a base that is attached to a distinct detectable label via a linker, said linker being cleavable with a water-soluble phosphine and said cleavable linker comprising a disulfide; and
a nascent polynucleotide complementary to the target polynucleotide, one of said provided nucleotides being suitable for incorporation into said nascent polynucleotide;(b) incorporating the nucleotide suitable for incorporation into said nascent polynucleotide; (c) detecting the detectable label of the incorporated nucleotide to identify the incorporated nucleotide; and (d) cleaving the disulfide linker of the incorporated nucleotide with a water-soluble phosphine. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13)
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Specification