Two distinct populations observed on a homogenous cells - (Nov/28/2018 )
Hi all, I often get two populations from my results of electroporated cells compared to non-electroporated control (pic attached). Left is non-electroporated,, right is electroporated.
Electroporation conditions:
3 million cells, 5ug plasmid, exponential (250V, 950 uF, Ω infinite , cuvette 4mm ).
What could be the reason?
I don't think it's doublet as the fluoresence intensity of both population almost similar when measure individually.
Thanks.
Hi Thomson,
First, I have never run flow cytometry myself, but while you are waiting for a real expert, I would postulate that your post-electroporation cells are a mixture of cells with the plasmid, and cells that did not receive the plasmid. That would be 2 populations. I don't think you get 100% conversion on electroporation, do you? (My experience is more with bacteria than cultured cells.)
As a control you could always try a dummy electroporation, with no plasmid added.
good luck!