Hi all,
I'm working on preparation of a stable cell line using hygromycin selection process.
I'm using HEK293 cell line and 200 ug /mL of hygromycin-containing media. However, I found that all cells dead maybe due to too high concentration of hygromycin for this system.
So I decreased the concentration of hygromycin, but I found that the cells were contaminated with bacteria which is resulted from too low concentration of hygromycin and can't prevent contamination from bacteria.
Therefore, I'm planning to use penicillin/streptomycin solution in a hygromycin selection process. In my opinion, the penicillin/streptomycin don't have big effect on the selection process but to confirm it, I'd like to get some opinion from you all guys.
Thank you so much in advance
Hi! Sure there is no problem in this, I've routinely used pen/strep solution and selected for different resistance markers (hygro, mtx)
Best, Eugene
- Most selection cassettes will incorporate Ampicillin resistance so this should work
Penicillin and streptomycin affect prokaryotes ( and not all of them) . They should have no effect on eukaryotic cells at standard concentrations. A good alternative is gentamycin which has a broad spectrum antibacterial effect. It’s also worth checking for problems with transfer of materials between your molecular biology lab and the culture hood as E. coli is very easily transferred unknowingly and will rapidly outgrow any eukaryotic cell in most media.
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