Recent developments in electrophoretic methods.

Abstract

13. Given the recent extended review by Vesterberg [J. Chromatogr., 480 (1989) 3-19] of electrokinetic methods, this survey has been restricted to the last decade, which has seen tremendous progress in several fields. DNA electrophoresis has experienced strong developments, both in the sequencing strategies (which have been largely automated with the use of fluorescent probes) and in pulsed field analysis of mega-DNA fragments, which has seen such developments as inverse-field, contour-clamped and rotating gel platforms, all allowing for straight band migration in each lane. Chromosome size mapping has now become a reality. Two-dimensional (2D) maps have also shown a dramatic improvement in performance, largely through the development of immobilized pH gradients, giving highly reproducible protein spots in the 2D plane and allowing the exploration of very narrow pH regions. Blotting techniques, combined with 2D mapping, allow sequence analysis and fingerprinting of a single polypeptide spot in a complex sample without resorting to lengthy chromatographic purification steps. Chromatophoresis generates a novel type of 2D mapping, based on hydrophobicity vs. size, rather than on charge vs. size, by direct coupling of a reversed-phase high-performance liquid chromatographic (HPLC) eluate to sodium dodecyl sulphate electrophoresis. The new rising star, capillary zone electrophoresis, offers speed, a large number of theoretical plates, selectivity and small sample requirements in a highly automated equipment.