Hi, I'm trying to co-transfect a mammalian cell line with two plasmids 1) a luciferase reporter plasmid and 2) a GFP-containing plasmid.
The luciferase plasmid contains a promoter of interest upstream and the GFP plasmid contains a mutated gene that I want to express. I want to see if the mutation from the GFP-plasmid affects the expression level of the gene in the luciferase plasmid. How do I confirm that my cells have successfully co-transfected both?
Thanks in advance for your help!
Hello Fidelia,
If I was in your place, I would check what is the emission wave length for the luciferase plasmid. If the two plasmids emit in different wave lengths, then I would transiently transfect the cells with both plasmids and run them through FACs. The FACs will allow you to distinguish between the cells with single and double transfection, and will give you information about the transformation efficiency.
hope that helps!
Good luck with your experiment
Olga
Hello,
Ibrahim, the western blot will give you informations about the expressed proteins, but it won't give you information about the efficiency of the double transformation.
Best
Olga
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