Transfection - Plasmid Uptake Control - (Jan/06/2010 )
I'm transfecting HEK 293T cells by nucleofection and I want a control for the amount of plasmid taken up in each transfection.
I've read that this can be done by Southern blot but isn't very quantitative and is an out-dated technique.
The alternatives I've come across are cotransfections with reporter plasmids, but surely plasmid-reporter plasmid interactions and different capactity for uptake between reporter and construct could skew results?
I was wondering what is generally done in the field?
Thanks for your time,
What about quantitative PCR with a standard curve?
You mean to check the efficiency of the electroporation, right? ( I dont know if I got it right).
You can use a plasmid producing gfp and just check the amount of green cells (IF or cytometry)