Hi all, I am transfecting Drosophila S2 cells for Luciferase assay. My cells are in 24 well plates, in 1ml S2 complete media (10% FBS + antibiotics). Is it okay if I just add DNA-Lipid complex (50ul) to cells directly ? Or do I need to wash and replace the cells with 1ml serum free media before adding the DNA-Lipid complex ?
(since Invitorgen claims transfection works with media containing serum and antibiotics with Lipofectamine 3000)
And also for S2 cells which one is better ? Lipofectamine 2000 or 3000 ?
I would avoid using Serum and treatment with antibiotics before/during lipofection.
I would propose alternatively to use electroporation (via AMAXA)
Well firstly, you shouldn't just add the Lipid-DNA mix as you need to measure the density of your cells so that you can plate them at the density you want. Therefore you need to resuspend the cells in fresh growth media and plate those.
Also the serum free media is part of the Lipid-DNA reaction so you will use both and the serum free media will roughly be 10% of the well.
When I did luciferase assays I used Graces insect medium, as my serum free media. Also I would recommend using mirus transit insect transfection reagent instead of the standard lipofection kits as it has been maximised for plasmid transfection into S2 cells and works with media containing serum. With that kit I always got a high transfection efficiency.
Good luck
- Badges
- Science method