I am doing a gene editing experiment and I want to edit two sites at the same time, so there are two resistance genes, can I add two antibiotics to the cell after transfection 72 hours or do I add one antibiotic to the cell first and then add the other antibiotic to the screen? Is it possible that screening after two weeks of transfection has no effect?
To my experience, researchers have used two antibiotics screening at the same time. It can either be first incorporation on only one antibiotics followed by selection and then second incorporation, or else incorporating different antibiotics at different sites at the same time.
You can use two antibiotics at the same time although depending upon the antibiotics there is some synergistic stress incurred by the cells due to having two drugs. So just be aware.
However do you need to both edits at the same time? It might be easier to do them sequentially if possible? Because depending upon efficiency of your experiments the frequency of getting both is certainly going to be lower than getting one.
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