Transient Transfection of HeLa cells. - (Feb/04/2008 )
Hi, I m new to transfection.
I now plan to use Lipofectamine 2000 as reagent to transient transfect HeLa cells in 24-well plate.
Can anyone kindly give me some comments on my below amount of plasmid used (total DNA used in each r'x = 800 ng):
Control 1
pGL3 empty vector: 300 ng
junk vector: 300 ng
beta-galactosidase normalizing vector: 200 ng
Control 2
pGL3 empty vector: 300 ng
Transcription factor construct: 300 ng
beta-galactosidase normalizing vector: 200 ng
Samples:
pGL3-basic (with promoter inserted): 300 ng
Transcription factor construct: 300 ng
beta-galactosidase normalizing vector: 200 ng
In each well I m gonna seed 8*104 cells in 500uL medium
Thank you in advance.
Have you optimized the transfection, like with a vector expressing fluorescent protein? If so, then go ahead.