Capacitively-induced pulsed-field gel electrophoresis: a novel method for DNA separation.
Present instruments used in pulsed field and conventional gel electrophoresis, encounter a number of real and conceived difficulties such as electrical hazard, ground leak current, electrical noises, formation of gas bubbles at the metal electrodes and production of Joule heat in the buffer-gel system. To overcome the above-mentioned problems a novel electrophoresis unit based on capacitively-induced pulsed field was designed and tested in which the applied high voltage is decoupled from the electrolyte (buffer-gel system). In order to achieve a higher performance, the primary pulse generator which has been fabricated to apply for capacitively-induced pulsed field electrophoresis was equipped with a modulating frequency pulse generator which produced a combination of low and high frequency waves. The newly designed electrophoresis unit was able to resolve the Lambda DNA fragments so that seven bands with an acceptable resolution were observed. By increasing the run time both the depletion of molecules from the wells and the resolution of the bands improved compare to the patterns obtained via conventional electrophoresis.
Ghourchian
Hedayatollah
H
Laboratory of Microanalysis, Institute of Biochemistry & Biophysics, University of Tehran, P.O. Box 13145-1384, Tehran, Iran. hadi@ibb.ut.ac.ir
Elyasvandi
Hossein
H
D002021:Buffers; D004247:DNA / Q000032:analysis* / Q000737:chemistry; D004573:Electrolytes / Q000494:pharmacology; D004586:Electrophoresis; D016521:Electrophoresis, Gel, Pulsed-Field / Q000295:instrumentation* / Q000379:methods*; D008970:Molecular Weight
D002021:Buffers; D004573:Electrolytes; D004247:DNA