Effectene Transfection Reagent (1 ml)

QIAGEN offers a non-liposomal lipid transfection reagent designed for efficient DNA transfection, with particular strength in primary cells and sensitive cell lines. Used together with a DNA-condensing Enhancer and Buffer EC, the formulation achieves high transfection efficiencies in eukaryotic cells, including insect cell lines such as S2 and Sf9, with minimal cytotoxicity. Both transient and stable transfection are supported, and the procedure can be carried out in the presence of serum and antibiotics using only low amounts of DNA.

The protocol is a two-step process. In the first step, DNA is condensed by the Enhancer in Buffer EC for 2–5 minutes. The reagent is then added and the mixture incubated for 5–10 minutes to allow complex formation. The resulting complexes are mixed with growth medium and added directly to the cells, with no requirement for complex removal. For adherent cells, optimal confluency at the time of complex addition is normally 40–80%. Each 1 ml of reagent supports approximately 160 transfections in 12-well plates.

Several parameters can be optimized to improve results: the reagent-to-DNA ratio, the overall amount of transfection complex, incubation time for gene expression, vector features such as promoter, origin of replication and plasmid size, cell density, and DNA quality. For endotoxin-sensitive cell lines and primary cells, DNA purified with EndoFree Plasmid Kits is recommended.