Posted 05 June 2002 - 03:48 AM
Posted 08 June 2002 - 06:40 AM
Posted 27 August 2004 - 04:20 AM
i had done stable trasnfection of SP2/0 cell line. After 48 hr i had added G418 at 200mug/ml, what is the maximum concentration i can use to select clone.
When should i change the media of cells and after how many days i shall serially dilute them?
Can anybody help me out, please?
Posted 29 August 2004 - 08:27 PM
resistant cells will continue growing and dividing in the presence of antibiotic, and you will get more and more dead cells with time in sensitive cell lines- I do my curves over 14 days. If you are trying to pick a concentration to select stable clones with- I normally go for the one that gives you dramatic cell death by about day 5, and no live cells at all by day 14.
As far as Shilpi's question goes: I'm not familiar with your particular cell line, but cell lines differ in their susceptibility to geneticin, and for some of my cells I have gone up to 1.2mg/ml. Change media as needed, as you would in normal tissue culture.