The mobility minima in pulsed-field capillary electrophoresis of large DNA.
Pulsed-field capillary electrophoresis represents a new tool for rapid and highly efficient separations of large biopolymers. The method has been utilized here to study dependencies of the electrophoretic mobility upon the frequency and pulse shape of applied voltage for large, double-stranded DNA molecules (5-100 kb) migrating in neutral polymer solutions. Two different shapes of alternating electric field (sine- and square-wave impulses) were examined with the frequency values ranging from 1 to 30 Hz. The linear dependence between duration of the forward pulse (at which the DNA molecule experiences a minimum mobility) and the product N.In(N) (where N is the number of base pairs) was experienced in field-inversion gel electrophoresis, while exponential dependence was found with the sinusoidal electric field. The mobility minima were lower in field-inversion electrophoresis than with the biased sinusoidal-field technique. The DNA (5 kb concatamers) was adequately separated using a ramp of frequency in the square-wave electric field, in approximately 1 h. The migration order of DNA fragments was referenced through adding a monodisperse DNA (48.5 kb) into the sample. The band inversion phenomena were not observed under any experimental conditions used in this work.
Sudor
J
J
Department of Chemistry, Indiana University, Bloomington, IN 47405, USA.
Novotny
M
M
D010582:Bacteriophage lambda / Q000235:genetics; D002197:Capillary Action; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D004279:DNA, Viral / Q000032:analysis / Q000737:chemistry*; D004560:Electricity; D004586:Electrophoresis / Q000379:methods*; D009690:Nucleic Acid Conformation
D004279:DNA, Viral