Invitrogen have added a new product, Lipofectamine™ LTX, to their extensive range of transfection products. Following on from other products in the Lipofectamine range, Lipofectamine™ LTX is a lipid based reagent for transfecting DNA and vector based RNAi into eukaryotic cells.
Lipofectamine™ LTX is an improvement on other Invitrogen transfection reagents as the protocol has been simplified into a simple single tube method that can be scaled up or down according to the size of culture plate in use. Briefly, DNA is diluted in an appropriate amount of cell media. The next step is to mix and add a small amount of Lipofectamine™ LTX. The mixture is incubated for 30 minutes (longer than other reagents) to allow complexes to form, before being added to the cells. The transfection reagent does not need to be removed from the cells, but can be replaced after 6 hours if required.
According to the manufacturers, this transfection reagent provides superior delivery and performance with less optimization in both adherent and suspension cells. In particular, this reagent has been optimized for hard to transfect cells lines such as primary cells and macrophages. This reagent can be used with PLUS™ Reagent, LINK TO PLUS REAGENT REVIEW another transfection reagent designed to enhance cationic lipid-mediated transfection. It should be noted that the manufacturers recommend using OPTI-MEM® I Reduced Serum medium for transfections. Other media may be used, but it is wise to run a preliminary test in case it affects the activity of the transfection reagent. Lipofectamine™ LTX may be used with or without serum which adds to its flexibility. It is worthwhile taking a look at the Invitrogen transfection website as the support offered is impressive. They have collected protocols for many cell lines and created an excellent reference database for the tissue culture scientist.
We have been trialing Lipofectamine™ LTX in our laboratory and have found it simple and very effective to use with no changes in protocol necessary to achieve high levels of transfection. The simplified protocol means we use less plasticware per transfection and need less transfection reagent per well. Cells that normally required a media change after about 6 hours survive better in this reagent, indicating reduced toxicity. We have transfected common mammalian cell lines (Vero, BHK-21 and PS-EK) according to the given protocol, and found an improvement in transfection efficiency in each case. We have only used the reagent with OPTI-MEM® I Reduced Serum medium.
Lipofectamine™ LTX Transfection Reagent From Invitrogen
Free of animal-origin components. Simple protocol and excellent support for different cell lines.
Longer incubation time required for DNA-reagent complexes.
The Bottom Line
An improvement on other Invitrogen products in efficiency and ease of use.