High-efficiency polyethylene glycol-mediated transformation of mammalian cells.
A new, high-efficiency method for transformation of mammalian cells with nucleic acids is described which yields 10(5)-10(6) plaques/micrograms poliovirus infectious RNA (iRNA). The optimized procedure consists of two steps: (1) exposure of cells to iRNA in a high ionic-strength buffer followed by (2) a brief exposure to a 35% polyethylene glycol (PEG) solution. Optimized conditions for each variable in the procedure are described. Under optimized conditions for PEG-mediated transformation with RNA, large numbers of transformants are recovered with plasmid DNA as well. The procedure presented is similar to other high-efficiency PEG-mediated methods previously described for the genetic transformation of both nonprotoplasted Escherichia coli and yeast.
D000818:Animals; D002460:Cell Line; D002472:Cell Transformation, Viral / Q000187:drug effects*; D006224:Cricetinae; D003412:Cricetulus; D005260:Female; D006367:HeLa Cells / Q000187:drug effects; D006801:Humans; D007700:Kinetics; D009994:Osmolar Concentration; D010053:Ovary; D010957:Plasmids; D017955:Poliovirus / Q000187:drug effects / Q000235:genetics*; D011092:Polyethylene Glycols / Q000494:pharmacology*; D010948:Viral Plaque Assay
D011092:Polyethylene Glycols