Cell growth inhibition by lipofectamine 2000 - (Aug/13/2008 )
I did an miRNA transfection experiment using lipofectamine 2000. The mock (lipofectamine 2000 only) showed cell growth inhibition than untreated, but I am not sure if there were cells dead because the supernatant was removed 24 hr post-transfection and only cells on the bottome were counted. I have two questions now:
1) Another person in the lab changed the transfection reagent 24 hr post-trasnfection, and he would like me to repeat his procedure since my experiment is to test his conclusion on another cell line. Any person did this before? Change the transfection reagent 24 hr later?
2) When I added the mock reagent, the lipofectamine solution had been "waiting" at room temperature for about 30 min, during the time I prepared for the lipofectamine and miRNA complex. Will this change the lipofectamine's conformation or something and harm the cells?
Thank you!
1) Another person in the lab changed the transfection reagent 24 hr post-trasnfection, and he would like me to repeat his procedure since my experiment is to test his conclusion on another cell line. Any person did this before? Change the transfection reagent 24 hr later?
2) When I added the mock reagent, the lipofectamine solution had been "waiting" at room temperature for about 30 min, during the time I prepared for the lipofectamine and miRNA complex. Will this change the lipofectamine's conformation or something and harm the cells?
Thank you!
Had the same or similar experience with lipofectamine... it is really toxic
to some cell lines 
... either change the media after 16-24 hours later (so that more cells survive) or use a better trfx reagent with lower cytotoxicity and less off-target effects...