TransIT® Transfection Reagents for DNA Mediated Virus Production

Rescue of Influenza Virus by Reverse Transfection With TransIT®-2020 Transfection Reagent. 293T and MDBK cells are reverse transfected with up to 10 separate influenza virus rescue plasmids per experiment. Reagent:total DNA complexes are prepared with TransIT®-2020 Transfection Reagent at a 3:1 ratio of reagent to total DNA. Virus-containing supernatants are harvested 48-96 hr post-transfection. Titers from transfected cells can be as high as 10⁷ pfu/ml. Rescued virus is then amplified on MDBK cells to increase titer and total yield. Viral stocks are subsequently titered by plaque assay on MDCK cells. Titers from representative rescue experiments with different viral strains and reassortants are shown. This experimental strategy is adapted from Neumann, et al. 2005 PNAS.

Data courtesy of Andrew Mehle, University of Wisconsin-Madison

TransIT®-293 Reagent Achieves High Transfection Efficiency in HEK 293 Cells. HEK 293 cells were transfected with pEGFP using TransIT®-293 Reagent in complete media. Cells were visualized under fluorescent microscopy 24 hours post-transfection.