transfection of shRNA into jurkat E61 cells - (Apr/13/2006 )
Hi...
I transfected jurkat E61 cells with
1. Pbabe Puro containing ShRNA
2. Pabe puro empty
....after one day I select with puromycin (1ug/ml and 3ug/ml).. I also select E61 cells (without any construct) with puromycin as a control..
Today is the 7th days since I put puromycin but I still can see many live cells in E61 cells (without any construct) plate (control) .... Is this normal?? because I read that it ussualy takes 5 days after puromycin for the the cells to die....
I saw colonies formed in Pbabe puro with shrna plate and in pbabe puro empty plate but I can't tell if that cells contain my construct because the control plate still have many live cells....
Can I know how to differentiate live and death cells for this suspended cells (jurkat T cells )?
Thank's alott