DEAE-dextran enhances electroporation of mammalian cells.
نویسندگان
چکیده
Electroporation has proven to be a convenient method for transfection of many cell types. One drawback has been that it requires large amounts of DNA, typically 20 to 80 /tg per transfection (1). Here we report that adding DEAE-dextran to the electroporation media dramatically increases electroporation efficiency in mammalian cells, lowering the amount of DNA required for efficient transfection. We routinely get efficient electroporation using 0.8 /tg DNA per transfection using the method described below. With luciferase activity as an indicator, electroporation efficiency is increased 7to 149-fold when DEAEdextran is added to the electroporation media (Table 1). Using plasmid recovery as the indicator in transient-transfections, 8to 14-fold more DNA is recovered from the cells when DEAEdextran is added to the electroporation media (Table 2). The basis for the enhanced electroporation efficiency is likely to be that DEAE-dextran causes adherence of DNA to the surface of cells (2). The resulting higher local concentration of DNA at the cell surface may result in enhanced uptake of DNA upon electroporation.
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ورودعنوان ژورنال:
- Nucleic acids research
دوره 20 24 شماره
صفحات -
تاریخ انتشار 1992