Sparsely cross-linked nanogels: a novel polymer structure for microchannel DNA sequencing
The present invention is generally directed to novel polymeric materials for use in the electrophoretic separation of nucleic acids. In particular, the novel polymer materials are sparsely crosslinked nanogels, dissolved in an aqueous buffer to form solutions with moderate to high viscosity. The present invention further provides methods for generating such novel polymers, and related methods of their use.
1. A polymeric nanogel composition comprising a sparsely cross-linked polymer structure, wherein said polymeric nanogel is formed from the polymerization of a water-soluble monomer used in the formation of polymers used for electrophoretic separation of nucleic acids, wherein said polymer comprises a water-soluble monomer cross-linked by from about 1×10 −8 mol % to about 1×10 −3 mol % non-ionic, hydrophilic cross-linking moiety as compared to the amount of water-soluble monomer present.
2. The composition of claim 1 , wherein said water-soluble monomer is selected from the group consisting of acrylamide, N,N-dimethylacrylamide (DMA), N,N-diethylacrylamide (DEA), ethylene oxide, N-hydroxy ethylacyrlamide, N-ethyoxyethylacrylamide, and N-methoxyethylacrylamide.
3. The composition of claim 1 , wherein said non-ionic, hydrophilic cross-linking moiety is selected from the group consisting of 1,4-butanediol diacrylate, 1,4-butanediol dimethacrylate, 1,3-butylene glycol diacrylate, 1,3-butylene glycol dimethacrylate, diethylene glycol diacrylate, diethylene glycol dimethacrylate, ethylene glycol diacrylate, ethylene glycol methacrylate, 1,6-hexanediol diacrylate, 1,6-hexanediol dimethylacrylate, neopentyl glycol dimethylacrylate, polyethylene glycol diacrylate, polyethylene glycol dimethylacrylate, tryethylene glycol diacrylate, tryethylene glycoldimethylacrylate, tripropylene glycol diacrylate, tetraethylene glycol diacrylate, tetraethylene glycol dimethylacrylate, divinyl esters of a polycarboxylic acid, diallyl esters of a polycarboxylic acid, diallyl maleate, diallyl fumarate, hexamthylenebismaleimide, divinyl adipate, diallyl succinate, a divinyl ether of ethylene glycol, cylcopentadiene diacrylate, N,N′-methylenebisacrylamide, N,N′-methylenebismethacrylamide, ethylene glycol dimethylacrylate and mixtures thereof.
4. The composition of claim 1 , wherein said hydrophilic cross-linking moiety is present in an amount selected from the group consisting of 1×10 −8 mol % as compared to amount of water-soluble monomer present, about 1×10 −7 mol % as compared to amount of water-soluble monomer present, about 1×10 −6 mol % as compared to amount of water-soluble monomer present, about 1×10 −5 mol % as compared to amount of water-soluble monomer present, about 1×10 −4 mol % as compared to amount of water-soluble monomer present, and about 1×10 −3 mol % as compared to amount of water-soluble monomer present.
5. The composition of claim 1 , wherein said nanogel composition has a matrix having low shear viscosity from about 300 centipoise to about 200,000 centipoise when said polymeric nanogel material is dissolved in an aqueous buffer.
6. The composition of claim 1 , wherein said polymer is formed using inverse emulsion polymerization.
7. A nanogel composition comprising a sparsely cross-linked polymeric material used for electrophoretic separation of nucleic acids formed from the polymerization of a water-soluble monomer mixture, wherein said water-soluble monomer mixture comprises at least two monomer agents, wherein the volume phase transition temperature of said nanogel composition is dependent upon the percent by weight of each of said at least two monomer agents in said water-soluble monomer mixture.
8. The nanogel composition of claim 7 , wherein said at least two monomer agents are selected from the group consisting of acrylamide, DMA, DEA, ethylene oxide, N-hydroxy ethylacyrlamide, N-ethyoxyethylacrylamide, and N-methoxyethylacrylamide.
9. The nanogel composition of claim 7 , wherein said at least two monomer agents are DEA and DMA.
10. The nanogel composition of claim 9 , wherein said monomer mixture is 40% wt DEA and 60% wt DMA.
11. The nanogel composition of claim 9 , wherein said monomer mixture is 50% wt DEA and 50% wt DMA.
12. The nanogel composition of claim 9 , wherein said monomer mixture is 60% wt DEA and 40% wt DMA.
13. The nanogel composition of claim 9 , wherein said monomer mixture is 70% wt DEA and 30% wt DMA.
14. The nanogel composition of claim 9 , wherein said monomer mixture is about 30% wt DEA and 70% wt DMA.