3 replies to this topic

[mcb56]

Hello, I have a stably transfected S2 cell line that has blasticidin resistance. Once the cells have been stably transfected, do I need to keep the blasticidin around to keep them trasnfected? I am not sure if the vector was integrated into the genome or not.

Thanks

[bob1]

It is a good idea to keep them under minimal selection (called maintenance) which is usually 1/2-1/10 of the normal selection level.  This is to ensure that the DNA is retained and not methylated or inactivated in some other fashion.

[MJD]

I agree with what Bob said, but in addition make sure to check your cells periodically i.e. every 3-6 weeks to make sure the cells have retained the vector and that the selecting agent is still active.  Also, you probably want to make freezebacks of the cells once you have stable transfectants, so if anything go wrong along the way you still have stably transfected S2 cells to go back to.  Cheers!

Edited by MJD, 11 September 2010 - 01:02 PM.

[krenny7]

keep an eye on the number of cells..Thats vey imporatant