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Methods Enzymol. 2013;529:209-26. doi: 10.1016/B978-0-12-418687-3.00017-3.

Generating mammalian stable cell lines by electroporation.

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  • 1Johns Hopkins University School of Medicine, Baltimore, MD, USA.


Expression of functional, recombinant mammalian proteins often requires expression in mammalian cells (see Single Cell Cloning of a Stable Mammalian Cell Line). If the expressed protein needs to be made frequently, it can be best to generate a stable cell line instead of performing repeated transient transfections into mammalian cells. Here, we describe a method to generate stable cell lines via electroporation followed by selection steps. This protocol will be limited to the CHO dhfr-Urlaub et al. (1983) and LEC1 cell lines, which in our experience perform the best with this method.

Copyright © 2013 Elsevier Inc. All rights reserved.


Colonies of cells; Dilute plasmid DNA; Electroporation; Methotrexate (MTX); Plating electroporated cells; Protocol

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