System for pH-neutral stable electrophoresis gel

• US 5,922,185 A
• Filed: 10/11/1996
• Issued: 07/13/1999
• Est. Priority Date: 03/31/1994
• Status: Expired due to Term

• Current Assignee: Life Technologies Corporation (Thermo Fisher Scientific Incorporated)
• Original Assignee: Novel Experimental Technology
• Inventors: Updyke, Timothy V., Engelhorn, Sheldon C.
• Primary Examiner(s): Beisner, William H.
• Assistant Examiner(s): NOGUEROLA, ALEXANDER STEPHAN
• Application Number:
  US08/730,678
• Time in Patent Office: 1,005 Days
• Field of Search: 204/456, 204/465, 204/466, 204/455, 204/450, 204/605, 204/606, 204/615, 204/468
• US Class Current: 204/468
• CPC Class Codes: G01N 27/44747 Composition of gel or of carrier mixture

Abstract

A gel and buffer system for gel electrophoresis wherein separation occurs at neutral pH.

Claims

1. Apparatus for use in a discontinuous buffer gel electrophoresis system, the apparatus comprising:

a slab of precast electrophoresis gel adapted for insertion in an electrophoresis container, the gel uniformly saturated with a gel buffer comprising a primary organic amine or substituted amine with a PKa near neutrality, titrated with hydrochloric acid or acetic acid, to a pH between pH 5.5 and pH 7.5.

2. The apparatus of claim 1, wherein the amine is Bis(2-hydroxyethyl) iminotris (hydroxymethyl) methane.

3. The apparatus of claim 1, wherein the electrophoresis gel is a polyacrylamide gel.

4. The apparatus of claim 1, wherein the acid is hydrochloric acid.

5. The apparatus of claim 4, wherein the electrophoresis gel is a polyacrylamide gel.

6. The apparatus of claim 1, wherein the acid is acetic acid.

7. The apparatus of claim 6, wherein the electrophoresis gel is a polyacrylamide gel.

8. The apparatus of claim 1, wherein the slab or precast gel is further adapted for fluid communication with a cathode buffer.

9. The apparatus of claim 8, wherein the cathode buffer comprises sulfite.

10. The apparatus of claim 8, wherein the cathode buffer comprises a thiol.

11. The apparatus of claim 8, wherein the cathode buffer comprises thioglycolic acid.

12. Apparatus for use in a continuous buffer gel electrophoresis system, the apparatus comprising:

a slab of precast electrophoresis gel adapted for insertion in an electrophoresis container, the gel uniformly saturated with a gel buffer comprising a primary organic amine or substituted amine with a pK_a near neutrality, titrated with a zwitterionic compound selected from the group consisting of N-tris(hydroxymethyl)methylglycine, piperazine-N, N'"'"'-2-ethanesuifonic acid, 3-(N-morpholino)-propanesulfonic acid, 2-(N-morpholino)-ethanesulfonic acid, N-(2-acetamido)-2-aminoethanesulfonic acid, 2-(N-morpholino)-2-hydroxypropanesulfonic acid, N-tris-(hydroxymethyl)-2-ethanesulfonic acid, N-2-hydroxyethyl-piperazine-N-2-ethanesulfonic acid, N,N-bis-(hydroxyethyl)-2-aminoethanesulfonic acid, and 3-(N-tris-(hydroxymethyl) methylamino)-2-hydroxypropanesulfonic acid, to a pH between pH 5.5 and pH 7.5.

13. The apparatus of claim 12, wherein the electrophoresis gel is a polyacrylamide gel.

14. The apparatus of claim 12, wherein the zwitterionic compound is N-tris(hydroxymethyl)methylglycine.

15. The apparatus of claim 12, wherein the amine is Bis(2-hydroxyethyl) iminotris (hydroxymethyl) methane.

16. The apparatus of claim 12, wherein the gel buffer comprises a compound selected from the group consisting of urea and formamide.

17. The apparatus of claim 12, wherein the slab or precast gel is further adapted for fiuid communication with a cathode buffer.

18. The apparatus of claim 17, wherein the cathode buffer comprises tris (hydroxy methyl) amino-methane.

19. The apparatus of claim 12, wherein the slab or precast gel is further adapted for fluid communication with an anode buffer.

20. The apparatus of claim 19, wherein the anode buffer comprises tris (hydroxy methyl) amino-methane.