Monstein H J, Geijer T
Biochem Int. 1986 Jun;12(6):889-96.
A simple, rapid and inexpensive scaled up miniprep procedure for preparing pure E. coli plasmid DNA is described. Bacterial cells were subjected to the boiling procedure and high molecular weight RNA was removed by LiCl-precipitation. Residual RNA and proteins were removed by subsequent treatment with RNase A and proteinase K/SDS respectively, followed by Sephadex G-50 and Sepharose 6B-Cl chromatography. The average yield from a 100 ml over-night bacterial suspension was 100 to 150 micrograms for pBR-322 DNA, and 250-500 micrograms for SP-6 derived recombinant plasmids. In addition, the described "scaled up" preparation does not require CsCl-ethidium bromide centrifugation; pure plasmid DNA can be prepared within 1 to 2 days.
本文描述了一种简单、快速且成本低廉的大规模小量制备法，用于制备纯大肠杆菌质粒DNA。细菌细胞经过煮沸处理，通过LiCl沉淀去除高分子量RNA。残留的RNA和蛋白质分别通过随后用RNase A和蛋白酶K/SDS处理去除，接着进行Sephadex G-50和Sepharose 6B-Cl柱层析。对于pBR-322 DNA，100 ml过夜细菌悬液的平均产量为100至150微克，对于SP-6衍生的重组质粒，产量为250 - 500微克。此外，所描述的“大规模”制备方法不需要CsCl-溴化乙锭离心；可在1至2天内制备出纯质粒DNA。
