Dry compositions and methods for gel electrophoresis
The invention provides dry compositions for preparing and loading a sample on a gel for electrophoretic separation. The dry compositions preferably include a tracking dye and a sedimenting agent selected from a five-carbon polyol (e.g., ribitol, arabitol, or xylitol),iso-erythritol, maltitol, and saccharine. Methods for making and using, as well as kits comprising the disclosed compositions, are also provided.
1. A dry composition for loading a sample on a gel, said composition comprising a tracking dye and a sedimenting agent, wherein said sedimenting agent is ribitol.
2. The composition of claim 1 , wherein the tracking dye is selected from the group consisting of Bromophenol Blue, Xylene Cyanol, Orange G, Cresol Red, Bromocresol Green, Ethidium Bromide, GelRed®, GelGreen®, Coomassie Brilliant Blue, Brilliant Cresyl Blue, Methylene Blue, Toluidine Blue, Azure A, Patent Blue VF, Acramine Yellow, Sudan III, Alizarin, Alizarine Blue, Alizarine Orange, Gallacetophenone, Hematoxylin, Scarlet Red, Galleon, Blue Dextran; Brilliant Green, Methyl Blue, Methyl Green, Bismark Brown Y, Bismark Brown R, Malachite Green, Neutral Red, Tolonium Chloride, Crystal Violet, and a combination thereof.
3. The composition of claim 2 , wherein the tracking dye is selected from the group consisting of Bromophenol Blue, Xylene Cyanol, Orange G, Cresol Red, and a combination thereof.
4. The composition of claim 1 , wherein the tracking dye is Bromophenol Blue.
5. The composition of claim 1 , further comprising a buffering agent and/or a chelating agent.
6. The composition of claim 5 , wherein the buffering agent is selected from the group consisting of tris(hydroxymethyl) aminomethane (Tris), phosphate buffered saline (PBS), 3-(N morpholino)propanesulfonic acid (MOPS), 2-(N morpholino)ethanesulfonic acid (MES), 4-(2 hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), and a combination thereof.
7. The composition of claim 5 , wherein the chelating agent is selected from the group consisting of ethylenediaminetetraacetic acid (EDTA), ethylene glycol tetraacetic acid (EGTA), diethylene triamine pentaacetic acid (DTPA), 1,2-bis(o-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid (BAPTA), and a combination thereof.
8. The composition of claim 5 , wherein the composition consists essentially of a tracking dye; a sedimenting agent, wherein said sedimenting agent is ribitol; a buffering agent; and a chelating agent.
9. The composition of claim 8 , wherein the tracking dye is Bromophenol Blue; the buffering agent is Tris; and the chelating agent is EDTA.
10. The composition of claim 1 , wherein the composition consists essentially of a tracking dye and a sedimenting agent; wherein said sedimenting agent is ribitol.
11. The composition of claim 10 , wherein the tracking dye is Bromophenol Blue.
12. A method of producing a sample for loading on a gel, said method comprising:
a) providing a dry composition comprising a tracking dye and a sedimenting agent, wherein said sedimenting agent is ribitol and b) contacting said composition with an aqueous solution, thereby producing said sample.
13. The method of claim 12 , further comprising mixing said composition with said aqueous solution.
14. The method of claim 12 , wherein the aqueous solution comprises a macromolecule selected from a nucleic acid, a peptide, and a protein.
15. The method of claim 14 , wherein the nucleic acid is selected from DNA and RNA.
16. The method of claim 12 , wherein the tracking dye is selected from the group consisting of Bromophenol Blue, Xylene Cyanol, Orange G, Cresol Red, Bromocresol Green, Ethidium Bromide, GelRed®, GelGreen®, Coomassie Brilliant Blue, Brilliant Cresyl Blue, Methylene Blue, Toluidine Blue, Azure A, Patent Blue VF, Acramine Yellow, Sudan III, Alizarin, Alizarine Blue, Alizarine Orange, Gallacetophenone, Hematoxylin, Scarlet Red, Galleon, Blue Dextran; Brilliant Green, Methyl Blue, Methyl Green, Bismark Brown Y, Bismark Brown R, Malachite Green, Neutral Red, Tolonium Chloride, Crystal Violet, and a combination thereof.
17. The method of claim 16 , wherein the tracking dye is selected from the group consisting of Bromophenol Blue, Xylene Cyanol, Orange G, Cresol Red, and a combination thereof.
18. The method of claim 12 , wherein the tracking dye is Bromophenol Blue.
19. The method of claim 12 , wherein the aqueous solution further comprises a buffering agent and/or a chelating agent.
20. The method of claim 19 , wherein the buffering agent is selected from the group consisting of Tris, PBS, MOPS, MES, HEPES, and a combination thereof.
21. The method of claim 19 , wherein the chelating agent is selected from the group consisting of EDTA, EGTA, DTPA, BAPTA, and a combination thereof.
22. The method of claim 19 , wherein the composition consists essentially of a tracking dye; a sedimenting agent, wherein the sedimenting agent is ribitol; a buffering agent; and a chelating agent.
23. The method of claim 22 , wherein the tracking dye is Bromophenol Blue; the buffering agent is Tris; and the chelating agent is EDTA.
24. The method of claim 12 , wherein the composition consists essentially of a tracking dye and a sedimenting agent, wherein the sedimenting agent is ribitol.
25. The method of claim 24 , wherein the tracking dye is Bromophenol Blue.
26. The method of claim 12 , wherein the aqueous solution comprises a buffering agent and/or a chelating agent.
27. The method of claim 26 , wherein the buffering agent is selected from the group consisting of Tris, PBS, MOPS, MES, HEPES, and a combination thereof.
28. The method of claim 27 , wherein the buffering agent is Tris present at a concentration of approximately 10 mM.
29. The method of claim 26 , wherein the chelating agent is selected from the group consisting of EDTA, EGTA, DTPA, BAPTA, and a combination thereof.
30. The method of claim 29 , wherein the chelating agent is EDTA present at a concentration of approximately 1 mM.
31. A dry composition for loading a sample on a gel, said composition comprising a tracking dye, which is Bromophenol Blue; a sedimenting agent, which is ribitol; a buffering agent, which is Tris; and a chelating agent, which is EDTA.