Hi,
I would like to co-transfect U2OS cells with GFP-containing plasmid, and my plasmid of interest using Polyjet transfection reagent. The main idea is to measure trasnfection efficiency, and that's the reason i'm using GFP plasmid.
I generally think that the "my plasmid"/GFP ratio should be >1, maybe something like 5 or 10 What do you think?
I usually trasnfect with 1000ng DNA.
Hello Oron,
I would use 1:1 equimolar ratio of both plasmid. If it is only for measuring transfection efficiency you can also set up a parallel transfection with only GFP plasmid to get an idea of the transfection efficiency.
If your aim is instead co-transfection (e.g. you will later sort GFP+ cells to enrich for cells carrying also your plasmid) you should definitely go for 1:1 ratio in the same transfection reaction. Please note that this will only enrich for your plasmid, and GFP+ cells will however not be 100% carrying also your plasmid.
Hope it helps
Cheers
Francesco Aulicino
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