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UNIVERSITY OF BUCHAREST
FACULTY OF PHYSICS Guest
2024-11-22 2:08 |
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Conference: Bucharest University Faculty of Physics 2004 Meeting
Section: Electricity and Biophysics
Title:
Large Size DNA Separation by Field Inversion Gel Electrophoresis
Authors:
Adriana Cristina Toma, Daniel Fologea
Affiliation:
Department of Electricity and Biophysics, Faculty of Physics, University of Bucharest
Faculty of Biology, University of Bucharest
E-mail
Keywords:
DNA analysis,chromosome,electrophoresis
Abstract:
Electrophoresis is one of the most important methods for DNA analysis, both for quantitative and qualitative estimations. Unfortunately, DNA molecules larger than 20 kpb (kilo base pairs) have the same mobility in the external electric field, so that these molecules will co-migrate to the same point. This drawback can be avoided using some special electophoresis technique, called Pulse Field Gel Electrophoresis (PFGE). In this work, a variant of this technique, Field Inversion Gel Electrophoresis (FIGE) is performed, in order to separate DNA molecules larger than 1 Mpb. In this variant of PFGE, the electric field is periodically switched to the opposite direction, according to the established time parameters. This is the most economical variant of PFGE, most of the electrophoresis components being provided by a classical electrophoresis system. The ramping time is established running a computer program that controls, also, the field inversion.
This technique was applied on yeast strain, allowing the observation of the chromosomes (number and size) up to 2 Mpb. This technique (electrocaryotyping) could be used for a direct analysis of chromosome by electrophoresis for a lot of strains
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