MECHANICAL DISRUPTION OF ESCHERICHIA-COLI FOR PLASMID RECOVERY

Five different mechanical cell disruption processes were evaluated as methods to extract plasmids from bacterial cells. The methods used wer e sonication, nebulization homogenization, microfluidization, and bead milling. The recovery yields of intact plasmids from the various meth ods were measured by quantitative gel electrophoresis. Bead milling an d microfluidization were found to have the highest potential for large scale extraction with total intact recoveries of over 90% and around 50%, respectively. Other methods resulted in substantial plasmid degra dation, with recoveries no greater than 20% of the total intact plasmi d. (C) 1995 John Wiley & Sons, Inc.