Pulleyblank D, Michalak M, Daisley S L, Glick R
Mol Biol Rep. 1983 Aug;9(3):191-5. doi: 10.1007/BF00775367.
A procedure is described for the isolation and purification of E. coli plasmid DNA by polyethylene glycol precipitation. The method is rapid, simple, inexpensive and amenable to both small and large scale manipulation. This procedure involves lysis of bacterial cells by treatment with pronase in sodium dodecyl sulfate, removal of chromosomal DNA by centrifugation, precipitation of residual nucleic acids with polyethylene glycol and removal of RNA by precipitation with LiCl. Plasmid DNA purified as described is pure enough for restriction endonuclease analysis, for use as a vector for the cloning of cDNA or synthetic DNA, or for use as a template in an E. coli transcription-translation cell-free system.
本文描述了一种通过聚乙二醇沉淀法分离和纯化大肠杆菌质粒DNA的方法。该方法快速、简单、成本低，适用于小规模和大规模操作。此方法包括用链霉蛋白酶在十二烷基硫酸钠中处理来裂解细菌细胞，通过离心去除染色体DNA，用聚乙二醇沉淀残留核酸，以及用氯化锂沉淀去除RNA。按所述方法纯化的质粒DNA纯度足以用于限制性内切酶分析、用作克隆cDNA或合成DNA的载体，或用作大肠杆菌无细胞转录-翻译系统的模板。
