checking transient transfection efficiency
Posted 23 March 2010 - 06:44 AM
I need to find a method that can be used to calculate transfection efficiency following my transient transfection. The plasmid I'm using does not contain a GFP site (sad) nor a site that will allow me to do a luciferase type assay, and because I'm doing this in suspension cells, doing a b-gal assay is also proving to be tough (unless someone knows of an alternative protocol that does not involve fixation).
Posted 23 March 2010 - 03:48 PM
Posted 24 March 2010 - 07:45 AM
How about a smear on a slide and then some IF for your protein of interest? Otherwise, do a parallel transfection with a GFP or other fluorescent plasmid and assume that the % transfected is similar.
I'm not sure what you mean by "some IF for your protein of interest". I had thought of doing say a co-transfection with a commercially-available GFP plasmid, except that I do not have a fluorescent microscope, currently. So I'm trying to do this assay using just a regular light microscope.
I do appreciate the thoughts... do you happen to have any more to deal with this problem?
Posted 24 March 2010 - 04:27 PM