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Qiagen Maxiprep - dissolve precipitated pellet

Cornelius Krasel krasel at wpxx02.toxi.uni-wuerzburg.de
Tue Jan 25 08:57:02 EST 2000


Student <mucineer at iname.com> wrote:
> Does anyone use Qiagen Maxiprep out there to prepare transfection grade
> DNA?

Sure :-)

> I have a problem and wonder if anyone can enlighten me on this. After the
> final IPA precipitation and EtOH wash, I dried the pellet by putting it in a
> 37 degree incubator briefly then tried to redissolve the pellet in
> 100microlitres of Tris pH8.5,

I wonder how people try to pellet from the rather large elution volume.
In this lab, we centrifuge the elution buffer in Corex glass vials as
recommended by Qiagen, don't wash the pellet but instead dissolve it in
300 ul water. The 300 ul of DNA solution is then transferred into a
Eppendorf vial and again precipitated with 2.5 vol EtOH. The pellet
generated from this precipitation is then dissolved in 100 ul H2O
(which usually takes quite a while), the concentration measured and
the volume adjusted if necessary.

BTW, the yield of the Qiagen columns increases dramatically in my
experience if you let the eluate sit in the coldroom overnight.

--Cornelius.

-- 
/* Cornelius Krasel, U Wuerzburg, Dept. of Pharmacology, Versbacher Str. 9 */
/* D-97078 Wuerzburg, Germany   email: phak004 at rzbox.uni-wuerzburg.de  SP4 */
/* "Science is the game we play with God to find out what His rules are."  */




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