从大肠杆菌中分离质粒高效化学转染人细胞培养物的简单技术

V. Manuvera, E. Grafskaia, V. Lazarev
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引用次数: 0

摘要

目前,市面上有大量用于分离高纯度质粒DNA的试剂盒,用于随后转染人类细胞系。然而,由于高成本和后勤问题,可能需要仅使用最简单的试剂和材料分离质粒DNA。我们提出了一种可能的方法,这种DNA分离,适合常规实验室使用。它基于众所周知的质粒DNA纯化原理和方法,成本最低,不需要特殊技能,并且易于扩展。该技术包括碱裂解、二氧化硅颗粒净化和凝胶过滤等步骤。结果表明,使用该方法分离的质粒转染人胚胎肾Expi293F细胞的效率并不低于使用Qiagen质粒maxi试剂盒(Qiagen, USA)纯化的质粒。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
A Simple Technique for Isolating Plasmids from Escherichia Coli for Efficient Chemical Transfection of Human Cell Culture
Currently, a large number of reagent kits are commercially available for the isolation of highly purified plasmid DNA for subsequent transfection of human cell lines. However, due to high cost and logistical problems, it may be necessary to isolate plasmid DNA using only the simplest reagents and materials. We present one of the possible methods for such DNA isolation, suitable for routine laboratory use. It is based on well-known principles and methods for plasmid DNA purification, has minimal cost, does not require special skills, and is easily scalable. The technique includes the steps of alkaline lysis, purification with silica particles and gel filtration. It was shown that plasmids isolated using the proposed method transfect human embryonic kidney Expi293F cells no less efficiently than plasmids purified using a specialized Qiagen plasmid maxi kit (�Qiagen�, USA).
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