Baum C, Forster P, Hegewisch-Becker S, Harbers K
Heinrich-Pette-Institut, Universität Hamburg, FRG.
Biotechniques. 1994 Dec;17(6):1058-62.
A number of transfection methods for mammalian cells are available; however, many cell lines may appear resistant to efficient transfection, or at best, necessitate lengthy optimization procedures in recommended protocols. We describe here an electroporation protocol that yields highly efficient gene transfer (20%-100% of surviving cells) in all 19 cell lines tested so far, including lymphoid, myeloid, glial, fibroblast and COS cells. Unlike earlier electroporation protocols, adaptation of this protocol to a cell line of interest only requires the optimization of a single parameter, i.e., the voltage, and can be performed within a day. Furthermore, it is also superior to transfection protocols for other methods (calcium phosphate precipitation, lipofection, DEAE-dextran transfection and transferrinfection) in terms of reproducibility, economy and average transfection efficiency for a wide variety of cell lines.
有多种用于哺乳动物细胞的转染方法；然而，许多细胞系可能对高效转染表现出抗性，或者充其量在推荐方案中需要冗长的优化程序。我们在此描述一种电穿孔方案，该方案在迄今为止测试的所有19种细胞系中都能产生高效的基因转移（20% - 100%的存活细胞），包括淋巴细胞、髓细胞、神经胶质细胞、成纤维细胞和COS细胞。与早期的电穿孔方案不同，将该方案应用于感兴趣的细胞系仅需优化单个参数，即电压，并且可以在一天内完成。此外，在可重复性、经济性以及对多种细胞系的平均转染效率方面，它也优于其他方法（磷酸钙沉淀法、脂质体转染法、DEAE - 葡聚糖转染法和转铁蛋白转染法）的转染方案。
