Transfection with MIN6 cells - problems with low counts (Apr/11/2005 )

Hello;

Would anyone be able to help-

I am trying to transfect MIN6 cells with my reporter plasmid and am not getting much of a reading above background. does anyone have any experience with this?

The transfections are in 96-well plates; cells are seeded 24hrs prior to transfection at a density of approx 1*10^4 cells/ well.
I use the lipofectamine reagent for transfection (have also tried FuGENE 6).
The plates were transfected for 48hrs before reading with the Dual-glo luciferase (Promega) reagent.

I normally transfect 100ng of my plasmid together with 12.5ng of the control Renilla pRL-sv40 plasmid.

If anyone out there has experience of transfecting MIN6 cells I would really appreciate some input as I am getting dismal readings!

thank you in advance

Your problem may be due to low transfection efficiency and/or toxicity.
My colleagues next door (endocrinology lab) routinely transfect insulinoma cells like RINm5F, INS-1 and pancreatic ductal line PANC-1 using GenCarrier-1. They are pretty satisfied with what they got so far.