McCoy Adam M, Collins Michelle L, Ugozzoli Luis A
Gene Expression Division, Bio-Rad Laboratories, Inc.
J Vis Exp. 2010 Jan 7(35):1662. doi: 10.3791/1662.
It is becoming increasingly apparent that electroporation is the most effective way to introduce plasmid DNA or siRNA into primary cells. The Gene Pulser MXcell electroporation system and Gene Pulser electroporation buffer (Bio-Rad) were specifically developed to easily transfect nucleic acids into mammalian cells and difficult-to-transfect cells, such as primary and stem cells. We will demonstrate how to perform a simple experiment to quickly identify the best electroporation conditions. We will demonstrate how to run several samples through a range of electroporation conditions so that an experiment can be conducted at the same time as optimization is performed. We will also show how optimal conditions identified using 96-well electroporation plates can be used with standard electroporation cuvettes, facilitating the switch from electroporation plates to electroporation cuvettes while maintaining the same electroporation efficiency. In the video, we will also discuss some of the key factors that can lead to the success or failure of electroporation experiments.
越来越明显的是，电穿孔是将质粒DNA或小干扰RNA（siRNA）导入原代细胞的最有效方法。基因脉冲器MXcell电穿孔系统和基因脉冲器电穿孔缓冲液（伯乐公司）是专门为将核酸轻松转染到哺乳动物细胞以及诸如原代细胞和干细胞等难以转染的细胞中而开发的。我们将演示如何进行一个简单的实验，以快速确定最佳电穿孔条件。我们将演示如何在一系列电穿孔条件下对多个样品进行操作，以便在进行优化的同时能够开展实验。我们还将展示如何将使用96孔电穿孔板确定的最佳条件应用于标准电穿孔比色皿，从而在保持相同电穿孔效率的情况下，便于从电穿孔板转换为电穿孔比色皿。在视频中，我们还将讨论一些可能导致电穿孔实验成功或失败的关键因素。
