The ability to introduce foreign DNA into eukaryotic cells, transfection, is critical to many aspects of modern biological research. Because of this, Targeting Systems has developed a line of transfection reagents, the Targefect reagents, that allows for high-efficiency transfection in most cell lines and some primary cells as well. We have used it in our lab in order to transfect cells to study gene regulation, to establish stably transfected cell lines and for retroviral packaging and production. We have found these reagents to be affordable and reliable as well as easy-to-use with low toxicity and a high transfection efficency.
The Targefect F1 reagent is a cationic lipid compound that has been tested in our lab in various cell lines as well as primary Kupfer cells. The primary advantage of this reagent is that it can be used with media that is supplemented with serum and antibiotics. The Targefect F2 reagent is a non-lipid cationic reagent that is used in medium without serum and antibiotics. Its primary advantage is that it has a component that allows for DNA condensation that enhances its uptake by cells. This can result in an increase in transfection efficiencies.
For the past five years, I have used these two products in my experiments and have found them to be reliable. The company supplies you with a protocol that can be used in many different types of cells, but I have found that I am able to get the highest transfection efficiencies with the lowest cellular toxicity when we use a 1:1ratio DNA to F1/F2 reagents. Thus, in a normal transfection reaction I use 2ug of DNA and 2ul of F1 or F2 reagent for each well in a 6 well dish. However, I have used as much as 10ul of F2 reagent on HEK293 cells with little toxicity. The DNA and F1 or F2 reagent are mixed in high glucose DMEM and incubated at 37C for as little as 20 minutes before adding it to the cells.
One other benefit to using these reagents is that Targeting Systems is a small company with a very responsive staff. They are easily accessible for any technical question you may have. The technical staff regularly communicates with me to make sure that I am 100% satisfied.
Sharon Yavrom
Graduate Student
University of Southern California