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One of the most useful tools in cell biology research is transfection, the introduction of foreign DNA into eukaryotic cells. In much of today's research, there is a growing need for the effective transfection of large quantities of cells. Polyplus-transfection's jetPEI transfection reagent is a highly efficient, low toxicity, water-soluble polymer that can be used in the presence of serum in culture media. Therefore, there is no need to change the culture medium before or after transfecting cells, making this method ideally suited for use with the Corning HYPERFlask cell culture vessel. This protocol was optimized using HeLa cells but has been successfully applied to a variety of cell types including Chinese hamster ovary (CHO) cells. This protocol is intended as a starting point that can be optimized by the end user for their cell lines.

This procedure describes plating cells into a HYPERFlask cell culture vessel and multiple wells of a 24 well plate. The 24 well plate will serve as a control for overall transfection efficiency as well as transfection efficiency of the large scale precipitate made for the HYPERFlask cell culture vessel. Once the procedure has been optimized for the HYPERFlask vessel, these samples are unnecessary. Should you choose to use a different size control well, scale your changes in reagents based on an equivalent mL/cm2.