Compositions for real-time, single molecule sequence determination

Abstract

A sequencing methodology is disclosed that allows a single DNA or RNA molecule or portion thereof to be sequenced directly and in substantially real time. The methodology involves engineering a polymerase and/or dNTPs with atomic and/or molecular tags that have a detectable property that is monitored by a detection system.

1 .- 22 . (canceled) 
     
     
         23 . A composition comprising:
 a labeled polymerase including two or more polymerase labels;   at least one labeled nucleotide or nucleotide analog including a nucleotide label; and   a target nucleic acid molecule;   wherein at least one of the labels is configured to emit a signal upon polymerization of the at least one labeled nucleotide or nucleotide analog by the polymerase.   
     
     
         24 . The composition of  claim 23 , wherein at least one of the fluorescent polymerase labels is associated with or covalently bonded to the polymerase. 
     
     
         25 . The composition of  claim 23 , wherein the nucleotide label is bonded to the β, γ or other terminal phosphate of the nucleotide or nucleotide analog. 
     
     
         26 . The composition of  claim 25 , wherein the nucleotide label of the at least one labeled nucleotide or nucleotide analog is configured to interact with at least one of the polymerase labels while the at least one labeled nucleotide or nucleotide analog is polymerized by the polymerase. 
     
     
         27 . The composition of  claim 26 , wherein at least one of the polymerase labels is a FRET donor, the nucleotide label is a FRET acceptor, and the interaction includes FRET. 
     
     
         28 . The composition of  claim 26 , wherein the composition includes at least two different types of labeled nucleotides or nucleotide analogs, each type including a different nucleotide label bound to the β, γ or other terminal phosphate of the labeled nucleotide or nucleotide analog. 
     
     
         29 . The composition of  claim 26 , wherein the composition includes at least three different types of labeled nucleotides or nucleotide analogs, each type including a different nucleotide label bound to the β, γ or other terminal phosphate of the labeled nucleotide or nucleotide analog. 
     
     
         30 . The composition of  claim 23 , further including a primer adapted to hybridize to a portion of the target nucleic acid molecule.