Hi all, my lab group is currently trying to transfect cells via electroporation, and our plasmid contains the blasticidin resistance gene to help select successfully transfected cells. What concentrations have you all had success with? I am seeing a bit of variety/range in the concentrations that can be used for this.
Hello Binh Duong,
The range is 2-20µg/ml. What works for me may not necessarily work for you. The best way would be to treat the un-transfected cells with a range of blasticidin concentrations to find the lowest concentration that completely kills the cells within a few days.
You may follow the below 5 steps.
1. Plate your un-transfected cells in standard culture conditions.
2. Create a series of blasticidin stock solutions at different concentrations, such as 0, 2, 5, 10, 15, 20µg/ml.
3. Add the different blasticidin concentrations to separate wells or dishes containing your cells.
4. Observe the cells over the next few days and note which concentrations effectively kill all the un-transfected cells.
5. Choose the lowest concentration of blasticidin that results in complete cell death within 3-5 days for your specific cell line.
It is always better to find the optimum blasticidin concentration yourself for your specific cell line by performing the above kill curve for blasticidin.
Best,
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