POLYNUCLEOTIDE SEQUENCING
A polynucleotide sequencing method includes a wash step that employs a composition including a polymerase. The composition may also include a plurality of nucleotides. The composition may be configured to prevent the polymerase from incorporating one of the plurality of nucleotides into a copy polynucleotide strand. The composition may be substantially free of Mg 2+
1 . A cartridge for use with a sequencing apparatus, the cartridge comprising:
(a) a first chamber comprising an incorporation composition comprising a first plurality of 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 second chamber comprising a detection composition for identifying the blocked, labeled nucleotide incorporated into the copy polynucleotide strand;
(c) a third chamber comprising a cleavage composition comprising a second plurality of reagents for removing a label and a blocking moiety from the blocked, labeled nucleotide incorporated into the copy polynucleotide strand; and
(d) a fourth chamber comprising a first wash composition comprising a polymerase and a plurality of nucleotides, wherein the first wash composition is configured to prevent the second polymerase from incorporating one of the plurality of nucleotides into the copy polynucleotide strand.
2 . The cartridge of claim 1 , wherein the first wash composition comprises a plurality of blocked nucleotides.
3 . The cartridge of claim 1 , wherein the first wash composition comprises a plurality of nucleotides labeled with a detectable moiety.
4 . The cartridge of claim 1 , wherein the first wash composition is substantially free of Mg 2+ .
5 . A kit for use with a sequencing apparatus, the kit comprising:
(a) a first container containing an incorporation composition comprising a first plurality of 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 second container comprising a detection composition for identifying the blocked, labeled nucleotide incorporated into the copy polynucleotide strand;
(c) a third container comprising a cleavage composition comprising a second plurality of reagents for removing a label and a blocking moiety from the blocked, labeled nucleotide incorporated into the copy polynucleotide strand; and
(d) a fourth container comprising a first wash composition comprising a polymerase and a plurality of nucleotides, wherein the first wash composition is configured to prevent the second polymerase from incorporating one of the plurality of nucleotides into the copy polynucleotide strand.
6 . The kit of claim 5 , wherein the first wash composition comprises a plurality of blocked nucleotides.
7 . The kit of claim 5 , wherein the first wash composition comprises a plurality of nucleotides labeled with a detectable moiety.
8 . The kit of any one of claims 5 , wherein the first wash composition is substantially free of Mg 2+ .