Hello everyone,
In a project I was working, we generated a construct containing Puromycin resistant gene. I want to transfect my GBM cells and select the cells with stable expression of the construct using Puromycin. Which concentration should I use and how long should I treat my cells with Puromycin?
Thanks in advance and sorry for the inconvenience,
Mervenur
Hi Mervenur,
The optimal concentration for a selection antibiotic depends on the robustness of you cells. You should definitely y apply a killing curve to find this optimum. For Puromycin a range of 10 – 100 µg/mL would be recommended. Chose a final concentration where after 10 days no viable calls can be detected anymore.
If you are working with cells a lot you should check-out Cellseeker (https://www.cellseeker.org/). This is an easy and free to use app to organize and inventory your cell collection online.
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