Schlaak Christina, Hoffmann Peter, May Karin, Weimann Andreas
Institut für Laboratoriumsmedizin und Pathobiochemie, Charité Universitätsmedizin Berlin, Campus Virchow-Klinikum Augustenburger Platz 1, D-13353, Berlin, Germany.
Biotechnol Lett. 2005 Jul;27(14):1003-5. doi: 10.1007/s10529-005-7867-z.
Recovery and purification of ligated plasmid DNA is a critical and limiting step in modern molecular cloning techniques prior to electroporation in E. coli. Four different DNA purification and desalting methods were compared in regard to simple handling, reproducibility, efficiency and cost-effectiveness using picogram and nanogram amounts of DNA. Microcolumn purification of DNA was up to two orders of magnitude more efficient compared to gel filtration, ethanol precipitation or drop dialysis.
在大肠杆菌中进行电穿孔之前,连接质粒DNA的回收和纯化是现代分子克隆技术中的关键且具有限制性的步骤。使用皮克和纳克量的DNA,就操作简便性、可重复性、效率和成本效益方面,对四种不同的DNA纯化和脱盐方法进行了比较。与凝胶过滤、乙醇沉淀或滴渗析相比,DNA的微柱纯化效率高出多达两个数量级。