High speed, high resolution compositions, methods and kits for capillary electrophoresis
View Patent ↗The invention provides compositions, methods and kits for high speed, high resolution of analytes by capillary electrophoresis starting with uncoated capillaries. The compositions comprise a sieving component, comprising a non-crosslinked acrylamide polymer, and a surface interaction component, comprising at least one uncharged and non-crosslinked water-soluble silica-adsorbing polymer. Methods for employing the novel compositions in capillary electrophoresis are provided. Kits comprising the novel compositions for use in the novel methods are also provided.
1. A composition for separating analytes by capillary electrophoresis comprising:
a sieving component comprising a non-crosslinked hydrophilic acrylamide polymer having a molecular weight (Mw) greater than about 3,000,000 Daltons (Da) and less than or equal to about 6,400,000 Da; and
a surface interaction component comprising one or more non-crosslinked hydrophobic polymers selected from the group consisting of poly(meth)acrylamide, N,N-disubstituted polyacrylamide and N-substituted polyacrylamide, wherein said N-substituents are selected from the group consisting of C 1 to C 3 alkyl, halo-substituted C 1 to C 3 alkyl, methoxy-substituted C 1 to C 3 alkyl, and hydroxyl-substituted C 1 to C 3 alkyl; wherein the sieving component and the surface interaction component differ in polymer chemical composition; and
wherein the composition does not include a crosslinked polymer gel.
2. The composition of claim 1 , wherein the one or more non-crosslinked polymers comprises poly(N,N-dimethylacrylamide) as a surface interaction component.
3. The composition of claim 1 , further comprising at least one denaturant wherein the denaturant is selected from the group consisting of at least one of formamide, urea, and 2-pyrollidinone.
4. The composition of claim 1 wherein the composition is configured to separate DNA fragments up to about 600 bases with a single base resolution of greater than about 0.58.
5. The composition of claim 1 , wherein the surface interaction component is present at a concentration in the range of about 0.001% to about 10% w/w.
6. A capillary electrophoresis element comprising:
an uncoated capillary;
a composition for separating analytes located within the uncoated capillary, the composition comprising: a sieving component comprising a uncrosslinked hydrophilic acrylamide polymer having a molecular weight (Mw) greater than about 3,000,000 Daltons (Da) and less than or equal to about 6,400,000 Da; and a surface interaction component;
wherein the surface interaction component comprises a solution of one or more non-crosslinked hydrophobic polymers;
the sieving component and the surface interaction component differ in polymer chemical composition; and
wherein the capillary electrophoresis element does not include a crosslinked polymeric gel.
7. The capillary electrophoresis element of claim 6 , wherein the one or more surface interaction component non-crosslinked hydrophobic polymers are selected from the group consisting of poly(meth)acrylamide, N,N-disubstituted polyacrylamide and N-substituted polyacrylamide, wherein said N-substituents are selected from the group consisting of C 1 to C 3 alkyl, halo-substituted C 1 to C 3 alkyl, methoxy-substituted C 1 to C 3 alkyl, and hydroxyl-substituted C 1 to C 3 alkyl.
8. The capillary electrophoresis element of claim 6 , wherein the surface interaction component non-crosslinked polymer is poly(N,N-dimethylacrylamide).
9. The capillary electrophoresis element of claim 6 , wherein the composition further comprising at least one denaturant wherein the denaturant is selected from the group consisting of at least one of formamide, urea, and 2-pyrollidinone.
10. The capillary electrophoresis element of claim 6 , wherein the uncoated capillary comprises silica, fused silica, quartz, silicate-based glass, phosphate glass, alumina-containing glass, or plastic channel.
11. The capillary element of claim 6 , wherein the surface interaction component is present at a concentration in the range of about 0.001% to about 10% w/w.
12. A kit for separating analytes by capillary electrophoresis comprising a composition comprising:
a sieving component comprising an non-crosslinked hydrophilic acrylamide polymer having a molecular weight (Mw) greater than about 3,000,000 Daltons (Da) and less than or equal to about 6,400,000 Da;
a surface interaction component comprising one or more non-crosslinked hydrophobic polymers selected from the group consisting of poly(meth)acrylamide, N,N-disubstituted polyacrylamide and N-substituted polyacrylamide, wherein said N-substituents are selected from the group consisting of C 1 to C 3 alkyl, halo-substituted C 1 to C 3 alkyl, methoxy-substituted C 1 to C 3 alkyl, and hydroxyl-substituted C 1 to C 3 alkyl;
the sieving component and the surface interaction component differ in polymer chemical composition; and wherein the composition does not include a crosslinked polymer gel.
13. The kit according to claim 12 , wherein the surface interaction component is poly(N,N-dimethylacrylamide).
14. The kit according to claim 12 , further comprising at least one denaturant selected from the group consisting of at least one of formamide, urea and 2-pyrollidinone.
15. The kit of claim 12 , wherein the surface interaction component is present at a concentration in the range of about 0.001% to about 10% w/w.