Effectene Transfection Reagent effectene转染试剂对各种类型的真核细胞DNA转染,尤其适用于原代细胞可在血清存在的情况下进行转染适合用于原代细胞或敏感的细胞系高转染效率而且较低的DNA使用量Low endotoxin level with <10 eu>
详细介绍
Effectene Transfection Reagent effectene转染试剂
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.
Effectene Transfection Reagent effectene转染试剂
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.
Images
Model of Effectene Principle High Transfection Efficiencies Using Effectene Reagent Transfection of Oligonucleotides Using Effectene Reagent Effectene Transfection Procedure Serum and DNA Quantity vs. Transfection Efficiency Effectene Reagent with Primary Cells
磷酸钙转染试剂是向哺乳动物细胞导入质粒DNA的最为常用的方法之一。由于其高度一致的pH值和盐浓度,我们的磷酸钙转染试剂可以保持一致的转染效率,而这是自行配置的转染试剂所不实现的。基于这个原因,CalPhos转染试剂成为了这15年以来广为流行的商品化磷酸钙转染试剂之一。另外,您可以关注Xfect Single Shots transfection reagent,这是一款使用方便的高性能转染试剂,冻干粉独立包装,只需要添加质粒DNA并用于您的细胞。