Betz J, Puchinger H, Träger L
Hoppe Seylers Z Physiol Chem. 1975 Mar;356(3):349-56.
Different methods for homogenization of cells of Streptomyces hydrogenans, for extraction of nucleic acids and for fractionation of the RNA and DNA obtained were critically examined. The only way to prepare high molecular weight rapidly labelled RNA and polysomes was to grind freeze-dried cells together with kieselguhr with a mortar and pestle. The best results for extraction of nucleic acids from the cell homogenate were obtained in the presence of diethyl pyrocarbonate (diethyl oxydiformate), yielding nucleic acids of considerable purity in a minimal amount of time. The best resolution of extracted nucleic acids was achieved by electrophoresis in 2% agarose acrylamide gels. This technique proved that during the cell homogenization and extraction procedure the bulk of nucliec acids was not degraded to low molecular weight material. An improved device for the registration of the profile of the absorption after gel electrophoresis is described.
对氢化链霉菌细胞匀浆、核酸提取以及所得RNA和DNA分级分离的不同方法进行了严格检验。制备高分子量快速标记RNA和多核糖体的唯一方法是用研钵和杵将冻干细胞与硅藻土一起研磨。在焦碳酸二乙酯(二乙氧基甲酰)存在的情况下,从细胞匀浆中提取核酸可获得最佳结果,能在最短时间内得到纯度相当高的核酸。通过在2%琼脂糖丙烯酰胺凝胶中进行电泳可实现对提取核酸的最佳分离。该技术证明,在细胞匀浆和提取过程中,大部分核酸不会降解为低分子量物质。描述了一种改进的用于记录凝胶电泳后吸收图谱的装置。