Time needed to determine G418 concentration for selection
Posted 13 December 2004 - 09:48 AM
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?
Posted 13 December 2004 - 11:15 PM
Posted 14 December 2004 - 08:40 AM
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!
Posted 25 September 2009 - 08:27 AM
I have set up a cell line 293, which was used for a stable transfection with pcDNA3.1/V5-HIS with a neomycin select gene. According to Invitrogen booklet, this antibiotics gene can be selected with Geneticin Selective reagent and a concentration of 100 to 1000 microgram/ml would be effective for the controll cells to die in 2-4 weeks. I used the Geneticin in 1000 microgram/ml concentration and now 5 weeks passed. Most of the control cells did not die even though last week I increased the concentration to 2000 microgram/ml. What is the problem? Can you give any advices on this matter?
Thank you very much.
Posted 26 September 2009 - 04:49 AM
Posted 10 November 2009 - 09:18 PM