Reversible terminator nucleotides and methods of use

Abstract

Disclosed herein a reversible terminator nucleotides and methods of use.

1 - 29 . (canceled) 
     
     
         30 . A polynucleotide in which the 3′ terminal nucleotide comprises a base and a sugar in which a label is attached to said base and a protecting group is attached to said sugar, wherein said label and said protecting group can be detached from said polynucleotide by an enzyme. 
     
     
         31 . The polynucleotide of  claim 30 , wherein said label and said protecting group can be detached from said polynucleotide under identical conditions. 
     
     
         32 . The polynucleotide of  claim 30 , wherein said label and said protecting group comprise a phosphate moiety. 
     
     
         33 . The polynucleotide of  claim 30 , wherein said label and said protecting group comprise a SO 3  moiety. 
     
     
         34 . The polynucleotide of  claim 30 , wherein said label and said protecting group comprise a C(O)R moiety. 
     
     
         35 . The polynucleotide of  claim 30 , wherein said enzyme is a phosphatase. 
     
     
         36 . The polynucleotide of  claim 35 , wherein said phosphatase is shrimp alkaline phosphatase. 
     
     
         37 . The polynucleotide of  claim 30 , wherein said enzyme is a sulfatase. 
     
     
         38 . The polynucleotide of  claim 30 , wherein said enzyme is an esterase. 
     
     
         39 . The polynucleotide of  claim 30 , wherein said label is a fluorescent label. 
     
     
         40 . The polynucleotide of  claim 30 , wherein said label is attached to said base by a fragmentable linker. 
     
     
         41 . The polynucleotide of  claim 40 , wherein said fragmentable linker comprises a trigger moiety selected from a group consisting of PO 4 , SO 3 , and C(O)R. 
     
     
         42 . The polynucleotide of  claim 41 , wherein said trigger moiety is a substrate for said enzyme. 
     
     
         43 - 51 . (canceled)