Hi, I need to transfect a melanoma cell line with a plasmid but I’m not sure how much DNA (ug) is necessary for the transfection and which plate is better for this, 96, 24 or 6-well plate.
I accept any advice, protocol and suggestions
Thanks!
I transfect MDA breast cancer cells with Lipofectamine 3000 and transfect about 1200-1600ng of plasmid DNA. My plasmid expresses really well so hence the possible lower end concentration. Try to follow the manufacture's instructions. I have only transfected in 6 well plates and it works fine. Also I use half of what they recommend for P3000 and it works just fine (but test it first).
We routinely perform transfections in our lab and have used the 6 well as well as 24 well format. Although 96 well should also be easy to use. Id suggest you to try 6-well plate transfection with 1.5- 2.0 ug of your favorite plasmid. If you trying to standardize transfection in its will be ideal to include a well for GFP transfection as a positive control which can be visualized next day itself. Also make sure you use high quality plasmid extraction method (midi prep) as mini-prep extraction have endotoxins, which significantly reduce transfection efficiency.
Goodluck!
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