High-efficiency transformation of bacterial cells by electroporation.
We have developed a method for efficiently generating transient pores in the outer membranes of Escherichia coli K-12 derivatives by using a new type of electroporation apparatus. The pores are large enough and persist long enough to facilitate the equilibration of plasmid molecules between the intracellular and extracellular spaces. The method has been used to transform bacterial cells with an efficiency greater than 10(9) transformants per microgram of plasmid. It has also been used to extract intact plasmid from transformed cells with efficiencies comparable to those of the traditional alkaline lysis or CsCl equilibrium density gradient techniques. The technique is simple and rapid, allowing a transformation or the preparation of microgram quantities of plasmid to be accomplished in minutes.
Calvin
N M
NM
Department of Biological Sciences, Stanford University, California 94305-5020.
Hanawalt
P C
PC
D002463:Cell Membrane Permeability; D002499:Centrifugation, Density Gradient; D004247:DNA / Q000032:analysis; D004560:Electricity / Q000295:instrumentation; D004574:Electromagnetic Fields; D055590:Electromagnetic Phenomena; D004926:Escherichia coli; D010957:Plasmids
D004247:DNA