transfection of mRNA

Paul Digard pd1 at mole.bio.cam.ac.uk
Sun Nov 19 10:54:26 EST 1995


> >We wish to transfect mRNA (synthesized in vitro) into a transformed 
rat cell
> >line and are seeking advice and suggestions from those who've had experience
> >doing this.

> We transfect in vitro transcriptions via electroporation, in RNAse-free PBS.
> The conditions will have to be worked out on your machine with your cells.
> We have calculated that ~60% of survivors (BHK-21 cells) take up the RNA.
> Also, lipofectin works well for RNA, albeit at lower efficiency than
> electroporation. Again, you'll have to optimize conditions. Note, lipofectamine
> was far less efficient than lipofectin for delivering RNA, although the reverse
> has generally been true for DNA in our lab. Just my thoughts...


We've used lipofectin (according to GIBCO's instructions) for 
transfecting b-gal mRNA into BHK cells, and have got up to around 30% 
positives.  Quite often less though....

Paul



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