Hi, I'm just looking for an optimal and reliable protocol for electroporation of T lymphoblasts with the Neon transfection system. We performed a previous experiment following the transfection settings from

"Transfection Optimization for Primary Human CD8+ Cells" J Immunol Methods. 2011 Sep 30; 372(1-2): 22–29. 

"Transfection of Human Naive CD4+ T cells with PHA Activation and Neon Electroporation" Bio-Protocol, Vol3, Iss15, Aug 05, 2013.

but our results showed no transfection: really low viability of T blasts, and no luciferase activity, nor GFP expression, which are the read-outs we are trying to use to monitorize transfection.

The setting we tried were 2200V, 12ms, 2 pulses.

T lymphoblasts were treated with anti-CD3 and PHA as activation controls, but we did'nt reach transfection at all.

Thank you in advance for your help,

Jordi