Purifying DNA from gel slices

Francois Coulier coulier at infobiogen.fr
Fri Nov 8 15:38:15 EST 1996


On Fri, 8 Nov 1996, David Bird wrote:

> Alison Reavley wrote:
> > 
> > Hi, I'm looking for a method of purifying plasmid DNA from gel slices
> > which does not involve phenol or any increase in temperature.  At the
> > moment I am trying electroelution but find that I am precipitating agarose
> > as well as DNA after the elution.  I need to get rid of this agarose again
> > without phenol or heating. I have also tried using Quiagen Qiaquick Gel
> > Extraction columns but have been frustrated by the small yield and I am
> > now using concentrations of DNA above those recommended in the protocol.
> > 
> > Any suggestions?
> > 
> > Alison Reavley
> > MRC Radiation and Genome Stability Unit,
> > Harwell,
> > Didcot UK.
> Many people in my department have used Glassmilk (pulverized glass
> beads) with success.  This preferentially binds DNA to glass beads, and
> the DNA is later eluted with a salt buffer, after washing with EtOH.  It
                                ^^^^^^^^^^^^
it supposed to be eluted in water, or, at most TE. Glassmilk works fine,
but again you need to dissolve agarose. Even with the help of sodium
iodide, you need to warm the sample to 55 degrees.

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Francois Coulier
INSERM Unite 119
27 bd Lei Roure
13009 Marseille
France

tel 33 (0) 4 91 75 84 11
fax 33 (0) 4 91 26 03 64)
email: coulier at infobiogen.fr
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