Enzymatic nucleic acid synthesis: methods for direct detection of tagged monomers

Abstract

Nucleotide triphosphate probes containing a molecular and/or atomic tag on a γ and/or β phosphate group and/or a base moiety having a detectable property are disclosed, and kits and method for using the tagged nucleotides in sequencing reactions and various assay. Also, phosphate and polyphosphate molecular fidelity altering agents are disclosed.

1 .- 54 . (canceled) 
     
     
         55 . A method for sequencing a nucleic acid molecule, comprising: (a) conducting a polymerase reaction in the presence of at least one detectably tagged nucleotide or nucleotide analog under conditions where the polymerase polymerizes at least one nucleotide or nucleotide analog, wherein the at least one detectably tagged nucleotide or nucleotide analog comprises a detectable tag bonded to a portion of the nucleotide or nucleotide analog that is released upon incorporation of the at least one nucleotide or nucleotide analog, and wherein the detectable tag alters fidelity of nucleotide or nucleotide analog incorporation by the polymerase. 
     
     
         56 . The method of  claim 55 , wherein the detectable tag is bonded to the β or γ phosphate of the nucleotide or nucleotide analog 
     
     
         57 . The method of  claim 55 , wherein the detectable tag is bonded to the β phosphate of the nucleotide or nucleotide analog. 
     
     
         58 . The method of  claim 55 , wherein the detectable tag is bonded to the γ phosphate of the nucleotide or nucleotide analog. 
     
     
         59 . The method of  claim 55 , wherein the detectable tag is ANS.