Principle

Effectene Transfection Reagent is an innovative non-liposomal lipid formulation that is used in conjunction with a special DNA-condensing enhancer and optimized buffer to achieve high transfection efficiencies. The enhancer first condenses the DNA molecules and Effectene Reagent subsequently coats them with cationic lipids （see figure "Model of Effectene Principle"）, providing a particularly efficient way of transferring DNA into eukaryotic cells. Effectene Reagent offers significant advantages over many liposome reagents and other transfection methods （1, 2）.

Procedure

The Effectene procedure has two steps. DNA is first mixed with Enhancer and a buffer that provides optimal salt conditions for efficient DNA condensation. This step requires just 2–5 minutes. Effectene Reagent is then added and the mixture is incubated for 5–10 minutes to allow Effectene–DNA complexes to form. The complexes are mixed with growth medium （which can contain serum and antibiotics）, and added directly to the cells. The cells are then incubated until harvested and analyzed for gene expression （see flowchart "Effectene Transfection Reagent"）.

Applications

Effectene Reagent is suitable for transient and stable transfection of a broad range of cell types （see figures "High Transfection Efficiencies Using Effectene Reagent" and "Transfection of Oligonucleotides Using Effectene Reagent"）. Because transfection can be performed in the presence of serum and requires low amounts of DNA （see figure "Serum and DNA Quantity vs. Transfection Efficiency"）, cytotoxicity is minimal. This makes Effectene Reagent particularly effective for primary cells （see figure "Effectene Reagent with Primary Cells"）. A searchable list of cell lines and primary cells successfully transfected using Effectene Reagent, as well as customer-developed transfection protocols, is available at the Transfection Tools web site — www.qiagen.com/TransfectionTools. View cell lines successfully transfected with this product.

High-throughput transfection

The application of recombinant DNA technology to fields such as drug discovery and development has led to an increased need for high-throughput transfection. Transfection using Effectene Reagent requires low amounts of DNA and minimal handling, and in addition, removal of transfection complexes is not required, making this reagent highly suitable for high-throughput screening. Effectene Reagent provides outstanding transfection efficiencies, excellent reproducibility, and minimal cytotoxicity in high-throughput transfection, and is available in bulk quantities.

Cited References

1. Ausubel, F.M., et al. eds. （1991） Current Protocols in Molecular Biology, Wiley Interscience, New York.
2. Sambrook, J., Fritsch, E.F., and Maniatis, T., eds. （1989） Molecular cloning — a laboratory manual, 2nd Ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor.