How long time needed to determine the concentration of G418? - (Dec/13/2004 )
I am trying to establish a stable transfection cell line. Before transfection, it is needed to determine a proper concentration of G418. As I know, a good concentration should kill all of the cell (non-transfected) within two weeks. Now I am trying the different concentration of G418 from 50-1ug/ml. The question is that how long time it is needed to determine the toxicity of the G418, if I have to wait two weeks?
IN general, G418 should kill all the cells in 4-5 days. At least, that has been the experience of the 3 labs I've been in. Usually a range of 500microgram - 1100 microgram /ml kills most cell lines within 5 days. Maybe 2 weeks works fine for you guys but you can be successful with 4-5 day toxicity and save alot of time. The clones usually become apparent within 2 weeks to 1 month.
Thank your advice. I agree with you, two weeks is a little long for me. I will firstly try to kill the cell within one weeks and get a good concentration. Have a nice christmas!