2 replies to this topic

[Oxford]

Hi,

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,

Adam

[madrius1]

What about quantitative PCR with a standard curve?

[laurequillo]

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)