Stable transfecion of BE2(M17) - (Jun/29/2004 )
Hi everybody
I am trying to find of the optimal G418 concentration for stable tarnsfection of BE2( M17) cells. After 12 days with 2 mg/ml G418 I have still alive cells. IIs it possible that my cell line is resistant to G418? Does anybody have exprience with that cells?
thanks in advance
Burcu
-anar123-
Hi!
Could your antibiotic be old? G418 has to be stored in the freezer and in the dark.
If it's ok, then try waiting another 2-3 days or increase concentration further(?).
Also, somewhat unlikely but possible, could your cells be contaminated by a G418 plasmid of some sort? It could happen if you have done many transfections before.
-kant0008-
QUOTE (kant0008 @ Jun 29 2004, 10:04 PM)
Hi!
Could your antibiotic be old? G418 has to be stored in the freezer and in the dark.
If it's ok, then try waiting another 2-3 days or increase concentration further(?).
Also, somewhat unlikely but possible, could your cells be contaminated by a G418 plasmid of some sort? It could happen if you have done many transfections before.
Could your antibiotic be old? G418 has to be stored in the freezer and in the dark.
If it's ok, then try waiting another 2-3 days or increase concentration further(?).
Also, somewhat unlikely but possible, could your cells be contaminated by a G418 plasmid of some sort? It could happen if you have done many transfections before.
Hi again
thanks for the answer. I tried G418 on Hela cells and worked perfectly. Indeed I did a PCR on M17 DNA for G418 casette and it is positive. We got these cells from somebody else. It was a good lesson: Check everything you received from others.
-anar123-