title: Generating mammalian stable cell lines by electroporation.
abstract: 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.
authors.affiliation: Johns Hopkins University School of Medicine, Baltimore, MD, USA.
authors.forename: Patti
authors.initials: P
authors.lastname: A Longo
authors.forename: Jennifer M
authors.initials: JM
authors.lastname: Kavran
authors.forename: Min-Sung
authors.initials: MS
authors.lastname: Kim
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Leahy
mesh_terms: D000818:Animals; D016466:CHO Cells; D002460:Cell Line / Q000166:cytology*; D006224:Cricetinae; D003412:Cricetulus; D018274:Electroporation / Q000379:methods*; D005822:Genetic Vectors; D008322:Mammals; D014162:Transfection / Q000379:methods*
keywords: Colonies of cells; Dilute plasmid DNA; Electroporation; Methotrexate (MTX); Plating electroporated cells; Protocol