how to set up a stable cell line? - why false positive always appears? (May/28/2006 )
I am to set up a stable cell line.
I have consulted several researchers around me, however, none of them has ever succeeded, with false positive cells appearing.
Had the cell mutated so that they were able to antagonize the selective substance such as G418?
G418 is a tough antibiotic that acts on dividing cell lines as it's a poison for ribozomes. Soslow or inactive cells remain very little affected by G418.
That's why G418 selection takes several weeks and you're not too sure of get rid of non transfected if you remove selection prssure too early. Moreover, in my lab we establih clones for G418 selection (1 cell needs to be resistant so the clone per se is resistant).
You need to increase the dose and/or keep selection to be sure.
Thank you very much for your help:) I will have a try~