Transfection Reagent (in vivo) is highly effective for DNA plasmid delivery in vivo, while not inducing an inflammatory response. TurboFect in vivo has been assessed for plasmid DNA delivery in both mice and rats and is suitable for DNA administration via various routes, including intravenous, intraperitoneal, and intratumoral injection. The amount of DNA and suggested injection volume will depend on the experimental animal and the route of administration as well as the targeted tissue or organ and the expression vector.
Transfection Reagent (in vivo) is a sterile solution of a cationic polymer in water. The polymer forms compact, stable, positively charged complexes with DNA. These complexes protect DNA from degradation and facilitate gene delivery in vivo. Reagent causes no detectable inflamatory response and is suitable for DNA administration via various routes, including intravenous, intraperitoneal, intratumoral injection, etc.
Transfection Reagent (in vivo) for 20 transfections using 50ug DNA.
Reagents Required but not Supplied:Sterile 5% glucose solution for dilution of DNA.
General Considerations:
DNA Quality Requirements:DNA quality is critical for successful transfection. An A260/A280 ratio of 1.8 or higher is recommended. DNA should be sterile and free of any contaminants such as endotoxins.
Choice of Promoter:High gene expression depends on both the promoter under which the gene of interest is expressed and the targeted tissue/organ. Cytomegalovirus (CMV) promoter is best known for high gene expression in a wide variety of cells. Some researchers prefer LTR, simian virus (SV40) or Rous sarcoma virus (RSV) promoters.
Transfection Reagent/DNA Ratio:The volume of transfection reagent used depends on the amount of DNA, transgene and animals to be transfected. The ratios presented in the protocols below are starting amounts and can be further optimized for best results.
Amount of DNA and suggested injection volume:The amount of DNA and suggested injection volume depend on the experimental animal and the route of administration (see protocol) as well as the targeted tissue or organ and the expression vector. To prevent precipitation of Transfection Reagent /DNA complexes, the final concentration of DNA in the injection mix should not exceed 0.5ug/ul.
Quality Control:Transfection efficiency was tested on BALB/c female mice (7-8 weeks old) using 50ug Firefly luciferase coding plasmid and 6ul of in vivo Transfection reagents per mouse. The delivery of the transgene was evaluated by measurement of Firefly luciferase activity in total protein extracted from the lungs of transfected animals. 100% of tested mice expressed the Firefly luciferase.
Storage and Stability:May be stored at 4°C. For long-term storage, aliquot and store at 4°C. Do not freeze. Aliquots are stable for 6 months after receipt. For maximum recovery of product, centrifuge the original vial prior to removing the cap.
