Electrophoretic Capture of Circular DNA in Gels - A Review
B. Akerman, Kenneth D. Cole
Results on electrophoretic capture of circular DNA in porous gels are reviewed. Processes, which cause arrest of circular forms of DNA during electrophoresis, can provide a very efficient separation mechanism for the purification of plasmids and bacterial artificial chromosomes, if the corresponding linear form is not trapped and therefore removed by the electric field. Two types of such topological traps have been proposed, impalement and lobster traps, and we here review the present experimental support for the existence of these two circle-specific mechanisms. We discuss experiments designed to characterize the traps in terms of the concentration of the traps as well as their efficiency and the capacity to trap both relaxed and supercoiled circular forms of DNA. Studies of the dynamics of the capture process showed that the average capture time was on the order of 10s at 20 V/cm, by which time the circles have migrated several hundred micrometers and have passed hundreds of traps. We also review results on attempts to improve the capacity and efficiency of the trapping process by modification of the gels either by enzymatic treatment or by cogelation of neutral polymers.