Nucleotide analogues

Abstract

The present invention provides methods, compositions, mixtures and kits utilizing deoxynucleoside triphosphates comprising a 3′-O position capped by a group comprising methylenedisulfide as a cleavable protecting group and a detectable label reversibly connected to the nucleobase of said deoxynucleoside. Such compounds provide new possibilities for future sequencing technologies, including but not limited to Sequencing by Synthesis.

1 .- 47 . (canceled) 
     
     
         48 . A composition comprising:
 i) at least one deoxynucleoside triphosphate according to the following structure:   
       
         
           
           
               
               
           
         
       
       wherein B is a nucleobase; and
 ii) at least one labeled deoxynucleoside triphosphate according to the following structure: 
 
       
         
           
           
               
               
           
         
       
       wherein D is selected from the group consisting of an azide, disulfide alkyl, disulfide substituted alkyl groups, disulfide allyl, and disulfide substituted allyl groups; B is a nucleobase; Linker comprises a cleavable oxymethylenedisulfide-containing site core, wherein said cleavable site core is selected from the group consisting of: 
       
         
           
           
               
               
           
         
       
       wherein R 1  and R 2  are independently selected alkyl groups; and Label is a label. 
     
     
         49 . The composition according to  claim 48 , wherein the nucleobase of i) or ii) is a non-natural nucleobase analog selected from the group consisting of 7-deaza guanine, 7-deaza adenine, 2-amino, 7-deaza adenine, and 2-amino adenine. 
     
     
         50 . The composition according to  claim 49 , wherein the nucleobase of i) and ii) is identical. 
     
     
         51 . The composition according to  claim 49 , wherein the nucleobase of i) and ii) is not identical. 
     
     
         52 . The composition according to  claim 48 , wherein said label is selected from the group consisting of fluorophore dyes, energy transfer dyes, mass-tags, biotin, and haptenes. 
     
     
         53 . The composition according to  claim 48 , wherein the composition further comprises a polymerase. 
     
     
         54 . The composition according to  claim 53 , wherein the composition further comprises a primer. 
     
     
         55 . The composition according to  claim 54 , wherein the primer is hybridized to a nucleic acid template. 
     
     
         56 . The composition according to  claim 55 , wherein said nucleic acid template is immobilized.