Journal article
Agarose electrophoresis of DNA in discontinuous buffers, using a horizontal slab apparatus and a buffer system with improved properties
Electrophoresis, Vol.14(1), pp.179-184
1993
Abstract
Using a horizontal slab apparatus with a buffer in the reservoirs at the level of the gel (“sea-level electrophoresis”), the retrograde discontinuous buffer system reported by Wiltfang et al. for sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of proteins was applied to DNA electrophoresis. This application yielded the advantages of an increased displacement rate of the moving boundary front and a decrease in the concentration of the counterion base in the resolving phase, which yielded reduced relative mobility values at equivalent gel concentrations and practicable low buffer concentrations. The change of relative mobilities (Rf) with a variation of field strength is decreased compared to that of the migration rate in the continuous Tris-boric-acid-EDTA (TBE) buffer and thus the robustness of the system is improved, as well as the efficiency of separation. The system of Wiltfang et al. has in common with previously described discontinuous DNA systems, that it is able to stack DNA from dilute samples and is insensitive to sample components with lower net mobilities than DNA, such as acetate. However, the variance of Rf at constant current density in the discontinuous buffer system is not improved over that of the migration rate at constant field strength in the continuous TBE buffer.
Details
- Title
- Agarose electrophoresis of DNA in discontinuous buffers, using a horizontal slab apparatus and a buffer system with improved properties
- Authors/Creators
- A. Zsolnai (Author/Creator)L. Orbán (Author/Creator)A. Chrambach (Author/Creator)
- Publication Details
- Electrophoresis, Vol.14(1), pp.179-184
- Publisher
- John Wiley & Sons Ltd
- Identifiers
- 991005544966607891
- Copyright
- © 1993 VCH Verlagsgesellschaft mbH
- Murdoch Affiliation
- Murdoch University
- Language
- English
- Resource Type
- Journal article
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- Collaboration types
- Domestic collaboration
- International collaboration
- Citation topics
- 2 Chemistry
- 2.166 Chromatography & Electrophoresis
- 2.166.730 Capillary Electrophoresis
- Web Of Science research areas
- Biochemical Research Methods
- Chemistry, Analytical
- ESI research areas
- Chemistry