Electrophoretic capture of circular DNA in gels.
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 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. Experiments designed to characterize the traps are discussed, regarding the concentration of the traps as well as their efficiency and capacity to trap both relaxed and supercoiled circular DNA. Studies of the dynamics of the capture process show that the average capture time is on the order of 10 s 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.
Akerman
Björn
B
Department of Physical Chemistry, Chalmers University of Technology, Kemivagen 10, S-412 96 Göteborg, Sweden. baa@phc.chalmers.se
Cole
Kenneth D
KD
D004270:DNA, Circular / Q000737:chemistry / Q000302:isolation & purification*; D004587:Electrophoresis, Agar Gel / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D005782:Gels / Q000737:chemistry; D009690:Nucleic Acid Conformation
D004270:DNA, Circular; D005782:Gels