( and primary cells we recommend using ...)Transfection Reagent Selector Kit Handbook

and primary cells, we recommend using DNA purified with EndoFree Plasmid Kits to ensure the highest transfection efficiencies. Reporter assay problem Include positive controls to ensure that the reporter assay is working properly. Excessive cell death Excessive exposure of cells to Transfection ReagentDNA complexes For Effectene: If sensitive cells (e.g. primary cells) or cell lines demonstrate extensive cell death after treatment with Effectene Reagent, remove the EffecteneDNA complexes after 618 hours. Wash cells carefully after removing complexes. With sensitive cell lines, we recommend 24 careful washing steps with complete medium rather than PBS.
For SuperFect: Most adherent cell lines yield optimal results when incubated with SuperFect DNA complexes for 23 h. If sensitive adherent cells (e.g. primary cells) or cell lines demonstrate extensive cell death after treatment with SuperFect reagent, reduce the exposure time of cells to complexes to 1 h. For sensitive suspension cells or cell lines, remove the complexes via centrifugation after a 23-hour incubation, and wash cells carefully. With sensitive cell lines we recommend 24 careful washing steps with complete medium rather than PBS. Amount of Transfection ReagentDNA complexes too high If cell death continues after reducing exposure times, decrease the amount of Transfection Reagent DNA complexes (see pipetting scheme ), but keep the ratio of Transfection Reagent to DNA constant.
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