I am working with HepG2 cells and transfecting them with a plasmid that upregulates the GABRA1 gene. I need to put them in selection media before I observe the results. HepG2 cells are resistant to Neomycin G418 (400 micrograms/mL), so I need to to make the selection media by putting the Neomycin in regular media. However, I don't know if the media I mix the Neomycin with should have Fetal Bovine Serum & Penstrep (which is what we usually culture cells in) or not.
During transfection, you should remove FBS and Penstrep because a lot of transfection reagents are sensitive to FBS. But after transfection, you should add in the FBS and Penstrep during your selection process.
Hello Harnoor Sachar
Your selection medium should contain Fetal Bovine Serum + 1% Penstrep + Neomycin. Penstrep should not have any effect on eukaryotic cells when used at optimal concentration.
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