Optimizing transfection conditions using LipoTAXI transfection reagent
Peter L. Pingerelli
Stratagene Cloning Systems
Clonetics Corporation, Walkersville, Maryland
LipoTAXI transfection reagent is a novel liposome reagent that offers low cellular toxicity and high transfection efficiencies for adherent and suspended cells. A procedure is described for optimizing the amounts of LipoTAXI transfection reagent and DNA used to transfect various cell types. Data from the transfection of four human cell strains from the Clonetics Corporation illustrate typical optimization results.
Many facets of gene analysis and recombinant protein expression rely on continuous development and improvement of transfection technology, the procedures for transferring foreign DNA into mammalian cells.1,2 LipoTAXI transfection reagent offers improved, state-of-the-art transfections for diverse cell types and can be used to transfect both adherent cells and cells growing in suspension. Generally, the two most important parameters for achieving optimal transfection efficiencies are the concentration of DNA and the volume of LipoTAXI transfection reagent. By measuring the signal intensity of a reporter gene and the cellular proteins across a matrix of conditions, researchers can easily optimize the amounts of DNA and LipoTAXI transfection reagent for any cell type.
The control plasmid provided with LipoTAXI transfection reagent encodes the
beta-galactosidase enzyme controlled by the cytomegalovirus (CMV) promoter.
Varying amounts of the reporter plasmid were mixed with different volumes of
LipoTAXI transfection reagent and added to cells plated in 12-well culture
dishes. For these optimization studies, Clonetics Corporation provided the
following cell types: small airway epithelial cell