MECHANICAL DISRUPTION OF ESCHERICHIA-COLI FOR PLASMID RECOVERY

被引:55
作者
CARLSON, A
SIGNS, M
LIERMANN, L
BOOR, R
JEM, KJ
机构
[1] PENN STATE UNIV,CTR BIOPROC RESOURCE,UNIVERSITY PK,PA
[2] APOLLON INC,MALVERN,PA
关键词
PLASMID RECOVERY; ESCHERICHIA COLI; CELL DISRUPTION;
D O I
10.1002/bit.260480403
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Five different mechanical cell disruption processes were evaluated as methods to extract plasmids from bacterial cells. The methods used were sonication, nebulization homogenization, microfluidization, and bead milling. The recovery yields of intact plasmids from the various methods were measured by quantitative gel electrophoresis. Bead milling and 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 degradation, with recoveries no greater than 20% of the total intact plasmid. (C) 1995 John Wiley & Sons, Inc.
引用
收藏
页码:303 / 315
页数:13
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