DNA prep for transient transfection

[Martin D. Leach]

In article <merle-0603951513020001 at 129.105.41.195>, merle ( ) wrote:

> > In Article <nick.5.2F559DCF at lmb1.rug.ac.be>
> > nick at lmb1.rug.ac.be writes:
> 
> > >
> > >As a coincidence, we had a discussion only yesterday about problems
with low 
> > >efficiencies of transient transfections in our lab.
> > >
> > >More precisely, transfections using the DEAE-dextrane method seem to
> give very 
> > >variable results, given one plasmid preparation to another.
> > >
> > >The low efficiency is independent of the person, the acceptor cells or
> the DNA 
> > >itself. In fact...the problem seems to be the preparation method
used. Sorry 
> > >to tell you so, but this all began when we started using QIAGEN columns!
> 

I find a slight batch to batch variation in transfectional efficiency (i
assume it is the efficiency) from Qiagen preps..Moreover, I sometimes find
a real difference when a new lot of qiagen columns are used...I had to
reduce how much bacteria (200ml vs. 500ml) on the columns so that I wasnt
overloading the newer columns...after this my transfections went back to
the higher value..

Martin

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