Field-inversion gel electrophoresis

• US 4,737,251 A
• Filed: 09/27/1985
• Issued: 04/12/1988
• Est. Priority Date: 09/27/1985
• Status: Expired due to Term

• Current Assignee: Washington University In St Louis
• Original Assignee: Washington University In St Louis
• Inventors: Olson, Maynard V., Carle, Georges F.
• Primary Examiner(s): Niebling, John F.
• Assistant Examiner(s): Starsiak, Jr., John S.
• Application Number:
  US06/781,283
• Time in Patent Office: 928 Days
• Field of Search: 204/182.8, 204/299 R
• US Class Current: 204/458
• CPC Class Codes: B01D 57/02 by electrophoresis treatment of water, waste water, sewage or sludge by electrophoresis C02F1/469; electrophoretic production of compounds or non-metals C25B7/00; investigating or analysing materials by using electrophoresis G01N27/26
• CPC Class Codes: G01N 27/44773 Multi-stage electrophoresis, e.g. two-dimensional electrophoresis

Abstract

Disclosed herein is a method and apparatus for gel electrophoresis which employs periodic inversion of the electric field essentially in one dimension. According to this system, which is defined herein as field-inversion gel electrophoresis, net migration is achieved by using a longer time or higher voltage in one direction than in the other direction.

Claims

1. An electrophoresis method to effect differential net migration, the extent of said migration being dependent on molecular size, of electrically charged macromolecular substances through a gel support in a single dimension, which method comprises:

subjecting electrically charged macromolecular substances selected from the group consisting of DNA, RNA, and detergent-protein complexes applied to a gel support to the repeated inversion of an electrical field along a single dimension, wherein the repeated inversion comprises cycles each consisting essentially of a first voltage in one direction of polarity for a first time interval, and a second voltage in the opposite direction of polarity for a second time interval,

wherein the migration effected by the total first voltages and first time intervals over all cycles does not equal the migration effected by the second voltages and second time intervals over all cycles, thus resulting in a net differential migration of said substances.

2. The method of claim 1 wherein each first voltage is equal to each second voltage and the total of the first time intervals is different from the total of the second time intervals.

3. The method of claim 1 wherein each first time interval is equal to each second time interval and each first voltage is different from each second voltage.

4. The method of claim 1 wherein for each successive cycle the first voltage bears the same ratio to the second voltage and the first time interval is equal to the second time interval, and wherein the magnitudes of said first and second voltages are monitonically greater or less in each successive cycle.

5. The method of claim 1 wherein for each successive cycle, the first time interval bears the same ratio to the second time interval, and the first voltage is equal to the second voltage, and wherein the first and second time intervals are monitonically greater or less in each succesive cycle.

6. The method of claim 1 wherein the electrophoresis gel support is an agarose gel.

7. The method of claim 1 wherein the substances are DNA molecules of about 15 kb or greater.

8. The method of claim 1 wherein the substances are applied at the same position in the gel support with respect to said single dimension.

9. The method of claim 8 wherein the substances form a mixture and said method results in a separation of the substances.

10. The method of claim 8 wherein the substances are applied next to each other at said position so as to migrate in parallel lines in the gel.

11. In a gel electrophoresis method to effect differential net migration of electrically charged marcomolecular substances through a gel support in a single dimension, wherein the net migration for each of said substances is a function of the size of said substance,

the improvement which comprises subjecting substances selected from the group consisting of DNA, RNA, and detergent-protein complexes, which have been applied to an electrophoresis gel support, to the repeated inversion of an electrical field along a single dimension, wherein the repeated inversion comprises cycles each consisting essentially of a first voltage in one direction of polarity for a first time interval and a second voltage in the opposite direction of polarity for a second time interval.

12. The method of claim 11 wherein each first voltage is equal to each second voltage and the total of the first time intervals is different from the total of the second time intervals.

13. The method of claim 11 wherein wherein each first time interval is equal to each second time interval and each first voltage is different from each second voltage.

14. The method of claim 11 wherein for each successive cycle the first voltage bears the same ratio to the second voltage and the first time interval is equal to the second time interval, and wherein the magnitudes of said first and second voltages are monitonically greater or less in each successive cycle.

15. The method of claim 11 wherein for each successive cycle, the first time interval bears the same ratio to the second time interval, and the first voltage is equal to the second voltage, and wherein the first and second time intervals are monitonically greater or less in each successive cycle.

16. The method of claim 11 wherein the electrophoresis gel support is an agarose gel.

17. The method of claim 11 wherein the substances are DNA molecules of about 15 kb or greater.

18. The method of claim 11 wherein the substances are applied at the same position in the gel support with respect to said single dimension.

19. The method of claim 18 wherein the substances form a mixture and said method results in a separation of the substances.

20. The method of claim 18 wherein the substances are applied next to each other at said position so as to migrate in parallel lines in the gel.