Multiple plasmid FuGENE 6 transfections

Ian A. York iayork at panix.com
Thu Sep 20 08:09:10 EST 2001


In article <3BA867FE.22154.1A7E6F at localhost>,
Wolfgang Schechinger <wolfsc at ibms.sinica.edu.tw> wrote:
>
>Keep in mind that if you have 50 (90) % transfection efficiency, on a very 
>rough calculation basis, your total Q will be .5 (.9) ^ 12 = 0.000244140625 
>(0.282429536481) if cells will take up 12 plasmids at all, assuming single 
>plasmid uptake events. And you probably will want to select for those cells 

I agree with your conclusion (that it's not going to work) but not with 
this point here.  In general in transfections plasmid uptake is not an 
independent event; cells which take up one plasmid are far more likely to 
take up many.  It's probably related to the nature of the cell (some cells 
in your dish are transfection-competent, some are not) and also to the 
transfection reagent--most transfection reagents bundle together many 
plasmids in the liposome (or whatever they use--FuGene is non-liposomal, 
but it's lipid-based and probably forms transfection complexes with 
multiple plasmids) so that all the plasmids are introduced into the same 
cell.

Ian 
-- 
    Ian York   (iayork at panix.com)  <http://www.panix.com/~iayork/>
    "-but as he was a York, I am rather inclined to suppose him a
     very respectable Man." -Jane Austen, The History of England




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