I am trying to transfect MDAM-468 breast cancer cells with GFP through electroporation.
I tried electroporating 50,000 cells in a 24-well plate with 2ug GFP and 1400 Volts, 10ms, 4 pulses (as per the the device Neon's protocol). I am getting a transfection efficiency of around ~20% (Instead of 80%). I then tried doing the same thing but with 5 million cells in a 10cm dish with 5ug GFP and I still got bad efficiency. I then tried a range of voltages from 1400-1800V and the best condition proved to be 1650V but the transfection efficiency was like ~30%.
Does anyone know the optimal conditions I should set on the Neon device to get a high transfection efficiency?
MDA-MB-468 is a breast cancer cell line, why are you trying to use electroporation? Electroporation is better suited for suspension cell lines (leukemia especially) and there it can have 80% efficiency. It isn't all that surprising that you're getting low transfection efficiency because MDA-MB-468 cells are generally transfected via polymers, liposomes, etc. We used this reagent (https://altogen.com/product/mda-mb-transfection-reagent-breast-adenocarcinoma/) for siRNA transfection into MDA-MB-468 and we got the 80%, but you're transfecting plasmids so the efficiency will end up being a bit lower. Good luck!
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