Transfections and GFP expression
martin.houle at REMOVE.drs.crchul.ulaval.ca
Mon Mar 1 12:12:21 EST 1999
In article <36D6734F.98E66BE0 at pet.mpin-koeln.mpg.de>, Mark Slack
<Mark.Slack at pet.mpin-koeln.mpg.de> wrote:
> We are doing lipofectamine transfections of a GFP expression plasmid
> into different cell lines all of which are done in duplicate. We do this
> by making a large cocktail of the DNA and lipofectamine mix and then
> pipette equal amount onto all cells.
> The problem we are having is that in one of the duplicates we may get
> really high transfection efficiency leading to high fluorescence and low
> background but the second duplicate we see low transfection and high
> background of fluorescence.
> I am looking for an explanation to this problem but do not think it is a
> mixing problem.
> I hope you can help.
> Many thanks.
Just so you know lipofectamine tends to fluoresce by itself. So that may
explain you high background. If at all possible, for you I would suggest
you use fugene 6 which does not give the high background that
lipofectamine does. You can check this out under a fluorescence microscope
and you'll see what I mean.
Martin Houle M.Sc.
Centre de Recherche du CHUL
Laval University, Quebec, Canada
More information about the Cellbiol