I am transfecting HUVEC cells with wild-type or point mutated forms of a gene that is cloned into pcDNA3.1. Post transfection, my cells are losing their adherence and, though I am not sure, I suspect that a significant proportion are dying.
I am using a jetPRIME transfection reagent designed for HUVEC cells according to suggested protocol.
This issue occurs when I use either transgene or vector alone, but not when I perform a mock transfection where no DNA is added.
The DNA is phenol/chloroform extracted so should be free of contaminates, so I think that the DNA itself may be harming the cells (I plan to do another phenol:chloroform extraction to be sure, though.
The plates are coated with 0.2% gelatin.
Not a lot of supervision here so I would very much appreciate any assistance.