Compositions, methods, kits, cartridges, and systems for sequencing reagents
The present disclosure relates to a composition including one or more modified nucleotide, wherein the modified nucleotide comprises a purine or pyrimidine base and a sugar moiety having a 3′-hydroxy blocking group, and a radical scavenger, wherein the composition is lyophilised. The present disclosure further relates to a composition including one or more functional protein; one or more functional protein activator; and one or more non-reducing sugar, wherein the composition is lyophilised. Also disclosed are methods of rehydration of one or more compositions described herein and kits including one or more compositions described herein. Further disclosed are cartridges including a flow cell comprising one or more reagent reservoirs, where the one or more reagent reservoirs include one or more compositions described herein.
1 . A composition comprising:
one or more nucleotides comprising a purine or pyrimidine base and a sugar moiety having a 3′-hydroxy blocking group, and
a radical scavenger, wherein the radical scavenger comprises gallic acid (GA), hydroxyethyl gallate (HEG), protocatechuic acid (PCA), catechin, or a combination thereof, and
wherein the one or more nucleotides and the radical scavenger are lyophilised together and nucleotide stability is increased by reducing lyophilisation-induced deblocking of nucleotides.
2 . The composition of claim 1 , further comprising:
one or more non-reducing sugars.
3 . The composition of claim 2 , wherein the one or more non-reducing sugars is trehalose, sucrose, inositol, maltodextrin, or dextran, mannitol, raffinose, cyclodextrin, melezitose, sorbitol, or a combination thereof.
4 . The composition of claim 2 , wherein the composition comprises at least about 5 wt. % non-reducing sugar.
5 . The composition of claim 2 , wherein the composition comprises about 5 wt. % or less non-reducing sugar.
6 . The composition of claim 1 , further comprising:
a polymerase.
7 . The composition of claim 1 , further comprising:
a functional protein activator.
8 . The composition of claim 7 , wherein the functional protein activator comprises magnesium, a magnesium salt, or combination thereof.
9 . The composition of claim 1 , wherein the 3′-hydroxy blocking group is a reversible blocking group.
10 . The composition of claim 1 , wherein the one or more nucleotides is linked to a detectable label.
11 . The composition of claim 10 , wherein the detectable label comprises a fluorophore.
12 . The composition of claim 1 , wherein the one or more nucleotides is linked to a detectable label via a cleavable linker.
13 . The composition of claim 1 , wherein the composition is a microsphere, a cake, or a combination thereof.
14 . The composition of claim 13 , wherein the composition has a cross-section of between about 0.1 mm and about 25 mm.
15 . The composition of claim 13 , wherein, when the composition is a microsphere, the microsphere is spherical, elliptical, or toroidal.
16 . The composition of claim 1 , wherein the composition comprises less than about 1 wt. % glycerol.
17 . The composition of claim 1 , wherein the composition comprises about 5 wt. % or less of a crowding agent.
18 . The composition of claim 17 , wherein the crowding agent comprises polyethylene glycol (PEG), polyvinyl pyrrolidone, dextran, maltodextrin, ficoll, polyacrylamide, or a combination thereof.
19 . The composition of claim 1 , further comprising a moisture content below about 10 wt. %.