An optimized electroporation protocol applicable to a wide range of cell lines.
An optimized electroporation protocol applicable to a wide range of cell lines.
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DOI：
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发表时间：
1994-12
期刊：
影响因子：
2.7
通讯作者：
C. Baum;P. Forster;S. Hegewisch-Becker;K. Harbers
中科院分区：
文献类型：
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作者：
C. Baum;P. Forster;S. Hegewisch-Becker;K. Harbers
许多哺乳动物细胞的转染方法是可用的;然而，许多细胞系可能对有效转染有抵抗力，或者最好的情况下，需要在推荐的方案中进行冗长的优化程序。我们在这里描述了一种电穿孔方案，该方案在迄今为止测试的所有19种细胞系中产生高效的基因转移（20%-100%的存活细胞），包括淋巴细胞、骨髓细胞、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，该方案对感兴趣的细胞系的适应仅需要优化单个参数，即，电压，并可以在一天内进行。此外，就多种细胞系的再现性、经济性和平均转染效率而言，它也上级其他方法的转染方案（磷酸钙沉淀、脂质体转染、DEAE-葡聚糖转染和转移感染）。
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.