Compositions for use in polyunucleotide sequencing

Abstract

A polynucleotide sequencing method comprises (i) removing a label and a blocking moiety from a blocked, labeled nucleotide incorporated into a copy polynucleotide strand that is complementary to at least a portion of a template polynucleotide strand; and (ii) washing the removed label and blocking moiety away from the copy strand with a wash solution comprising a first buffer comprising a scavenger compound. Removing the label and blocking moieties may comprise chemically removing the moieties. The first buffer may also comprise an antioxidant and may be used in a scanning buffer used during a nucleotide detection step.

1 - 9 . (canceled) 
     
     
         10 . A cartridge for use with a sequencing apparatus, the cartridge comprising a plurality of chambers containing a plurality of compositions, wherein each chamber contains a single composition, wherein the compositions consist of:
 (a) reagents for incorporating a blocked, labeled nucleotide into a copy polynucleotide strand complementary to at least a portion of a template polynucleotide strand;   (b) a buffer composition comprising an antioxidant and a scavenger;   (c) reagents for chemically removing a label and blocking moiety from the blocked, labeled nucleotide incorporated into the copy strand; and   (d) a wash buffer.   
     
     
         11 . The cartridge of  claim 10 , wherein the scavenger comprises a disulfide moiety. 
     
     
         12 . The cartridge of  claim 10 , wherein the scavenger is lipoic acid or 3,3′-dithiodipropionic acid (DDPA). 
     
     
         13 . A kit for use with a sequencing apparatus, the kit comprising a plurality of chambers containing a plurality of compositions, wherein each chamber contains a single composition, wherein the compositions consist of:
 (a) reagents for incorporating a blocked, labeled nucleotide into a copy polynucleotide strand complementary to at least a portion of a template polynucleotide strand;   (b) a buffer composition comprising an antioxidant and a scavenger;   (c) reagents for chemically removing a label and blocking moiety from the blocked, labeled nucleotide incorporated into the copy strand; and   (d) a wash buffer.   
     
     
         14 . The kit of  claim 13 , wherein the scavenger comprises a disulfide moiety. 
     
     
         15 . The kit of  claim 13 , wherein the scavenger is lipoic acid or 3,3′-dithiodipropionic acid (DDPA). 
     
     
         16 - 17 . (canceled) 
     
     
         18 . The kit of  claim 13 , wherein the scavenger comprises an azide moiety. 
     
     
         19 . The kit of  claim 13 , wherein the antioxidant is ascorbate, acetovanillone, or 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid. 
     
     
         20 . The cartridge of  claim 10 , wherein the scavenger comprises an azide moiety. 
     
     
         21 . The cartridge of  claim 10 , wherein the antioxidant is ascorbate, acetovanillone, or 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid.