Labelled nucleotides

Abstract

Nucleosides and nucleotides are disclosed that are linked to detectable labels via a cleavable linker group.

1 - 25 . (canceled) 
     
     
         26 . A nucleotide or nucleoside molecule, having a base that is linked to a detectable label via a cleavable linker, wherein the label and linker construct is of a size or structure sufficient to block the incorporation of a further nucleotide into a growing polynucleotide strand. 
     
     
         27 . The molecule of  claim 26 , wherein the base is a purine or a pyrimidine. 
     
     
         28 . The molecule of  claim 26 , wherein the base is a deazapurine. 
     
     
         29 . The molecule of  claim 26 , wherein the molecule is a deoxyribonucleotide triphosphate. 
     
     
         30 . The molecule of  claim 26 , wherein the detectable label is a fluorophore. 
     
     
         31 . The molecule of  claim 26 , wherein the linker is photocleavable. 
     
     
         32 . The molecule of  claim 31 , wherein the linker is based on O-nitrobenzyl. 
     
     
         33 . An oligonucleotide comprising the nucleotide molecule of  claim 26 . 
     
     
         34 . A method of labelling a nucleic acid molecule, the method comprising incorporating the nucleotide or nucleoside molecule of  claim 26  into the nucleic acid molecule. 
     
     
         35 . A method for determining the sequence of a target single stranded polynucleotide, comprising monitoring the sequential incorporation of nucleotides into a complementary strand of the target single stranded polynucleotide, wherein each of the nucleotides is a nucleotide according to  claim 26 , and wherein the identity of each nucleotide incorporated is determined by detection of the label linked to the base. 
     
     
         36 . The method according to  claim 35 , wherein said monitoring comprises:
 a) providing four types of nucleotides according to  claim 26 , wherein the detectable label linked to each type can be distinguished upon detection from the detectable label used for the other three types;   b) incorporating a nucleotide into the complementary strand of the target single stranded polynucleotide;   c) removing non-incorporated nucleotides;   d) detecting the label of the incorporated nucleotide of step b), thereby determining the type of nucleotide incorporated;   e) removing the label of the incorporated nucleotide of step b); and   f) repeating steps a)-e) one or more times, thereby determining the sequence of the target single stranded polynucleotide.   
     
     
         37 . The method according to  claim 35 , wherein the target single-stranded polynucleotide is immobilised on a solid support. 
     
     
         38 . A kit comprising:
 a) four individual nucleotides, wherein each nucleotide has a base that is linked to a detectable label via a cleavable linker, wherein the label and linker construct is of a size or structure sufficient to block the incorporation of a further nucleotide into a growing polynucleotide strand, and wherein the detectable label linked to each nucleotide can be distinguished upon detection from the detectable label used for the other three nucleotides; and   b) packaging materials therefor.   
     
     
         39 . The kit of  claim 38 , wherein two or more of the individual nucleotides are mixed. 
     
     
         40 . The kit of  claim 38 , wherein the base is a purine or a pyrimidine. 
     
     
         41 . The kit of  claim 38 , wherein the base is a deazapurine. 
     
     
         42 . The kit of  claim 38 , wherein the detectable label is a fluorophore. 
     
     
         43 . The kit of  claim 38 , wherein the linker is photocleavable. 
     
     
         44 . The kit of  claim 43 , wherein the linker is based on O-nitrobenzyl. 
     
     
         45 . The kit of  claim 38 , further comprising a polymerase and a buffer appropriate for the action of the polymerase.