The Influence Of Agarose Concentration In Gels On The Electrophoretic Trapping Of Circular Dna
| Autor: | Kenneth D. Cole, Björn Åkerman |
|---|---|
| Rok vydání: | 2003 |
| Předmět: |
Chromatography
Chemistry Process Chemistry and Technology General Chemical Engineering Direct current Analytical chemistry Filtration and Separation General Chemistry Trapping Circular DNA Electrophoresis chemistry.chemical_compound Field Inversion Gel Electrophoresis Electric field Kilobase pairs Agarose |
| Zdroj: | Separation Science and Technology. 38:2121-2136 |
| ISSN: | 1520-5754 0149-6395 |
| DOI: | 10.1081/ss-120021616 |
| Popis: | The density and structure of the electrophoretic traps for circular DNA in gels were strongly influenced by the concentration of agarose used to form the gels. Agarose gels were cast in a range of concentrations from 0.25% to 2.5% (1% is 1 g/100 mL). The trapping behaviors of two DNA circles with sizes of 13 kilobase pairs (kbp) and 52.5 were studied. The electric field strength (E) for the onset of trapping, the apparent trap density, and release characteristics of circles from traps were studied using both direct current and field inversion gel electrophoresis (FIGE) experiments. Direct current mobility indicated that the onset of trapping, indicated by a reduction in velocity and increase in band smearing, occurred at significantly lower value of E in 0.25% gels compared to 1% gels or 2% gels. The critical value of E for complete immobilization of the 13 open circular form was 16 V/cm in 0.25% gels, and 20 V/cm in 1% and 2% gels. FIGE mobility measurements, done by varying the reverse times, were used ... |
| Databáze: | OpenAIRE |
| Externí odkaz: |
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