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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