Chakrabarti A, Sitaric S, Ohi S
Department of Biochemistry, School of Medicine, Meharry Medical College, Nashville, Tennessee 37208.
Biotechnol Appl Biochem. 1992 Oct;16(2):211-5.
An expedient procedure for large-scale plasmid isolation from Escherichia coli strains without using ultracentrifugation or special setups or reagents is described. The protocol, which utilizes a modified alkaline extraction procedure as well as differential precipitations by isopropanol and lithium chloride, is simple and rapid and yet produces plasmid DNA with a yield of about 2 mg/liter culture. The isolated plasmids consisted of mostly monomeric and dimeric covalently closed circular DNA. The plasmids could be digested by various restriction endonucleases and were compatible with gene cloning, transfection-gene expression, and viral production.
本文描述了一种从大肠杆菌菌株中大规模分离质粒的简便方法，该方法无需使用超速离心、特殊装置或试剂。该方案采用改良的碱提取法以及异丙醇和氯化锂的分级沉淀法，操作简单、快速，且质粒DNA的产量约为2mg/升培养物。分离得到的质粒主要由单体和二聚体共价闭合环状DNA组成。这些质粒可用各种限制性内切酶消化，适用于基因克隆、转染-基因表达和病毒生产。
