Some-what large scale transfection - (Jan/08/2009 )
I'm attempting to scale-up my usual adherent CHO cell 96-well transfections to T-75 flasks and I need some help. Normally, I'm dealing with co-transfections: so I take my two plasmids and mix with 100ul of serum-free Ham's F-12K, then add about 6ul of Fugene HD, incubate, and then the resulting DNA: liposome complex is added to my cells.
How do I go about scaling this up to accommodate a T-75? What is the best/optimal cell density???
Read the Fugene manual, it has some very good information about seeding density and volumes of reagents to use.
Otherwise, work out the area of the wells that you transfect normally, then scale to the T-75 area.