Does anyone have a GOOD electroporation protocol for HeLa cells? I mean giving at least 90% transfection efficiency? I don't mind killing 50% of the cells if the remaining cells are all transfected!
you don 't need electroporation, I used fugene 6 Roche with 50% confluent cells (cells split the day before) and Optimem (Gibco), adding after 12-16 h 1 ml RPMI 10% serum (more o less). After 24h I get > 90% expession of a pGFP, as analysed by FACS.
Electroporation works better with non-adeherent cells, in my knowledge.
I used Xtremegene for transfection with very high efficiency. The transfection was done under serum free medium (optimem in both culture dish and for preparing the transfection mix), DNA 1ug, 20ul Xtremegene reagent.