I am trying to do transient co-transfections using HeLa cells and FuGENE 6 reagent. My luciferase reporter construct (in pGL3-basic) is co-transfected with the firefly control reporter driven by sv40. I do these is 96-well plates.
The problem is that I am not really getting any consistent readings even when i do exaclty the same protocol. I have checked DNA purity, for cell infection and all seems fine. I get very dismal numbers for the firefly readings and ok figures for the control renilla. Does anyone do 96-well transfections using HeLa cells that could help? how much DNA should i use/well? Currently i transfect 100ng DNA/well(+12.5ng control vector).
Any advice much appreciated
Edited by shojjahd, 01 March 2005 - 03:43 AM.