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Qiagen Maxiprep - dissolve precipitated pellet

CSC un691cs at genius.embnet.dkfz-heidelberg.de
Tue Jan 25 08:27:57 EST 2000


On Tue, 25 Jan 2000, Student wrote:

> Dear All
> 
> Does anyone use Qiagen Maxiprep out there to prepare transfection grade DNA?
> 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, however the pellet doesn't seem to dissolve
> very well. There are lots of white "bits" in the solution, it doesn't help
> when I increase the vol. to 160microlitres with warming to 37 degrees.

> Can anyone give a hint? THanks.

Just dissolve it, the white stuff has nothing to do with RNA. Probably
glycogen or something like that. After dissolving, vortex briefly, then
spin the white stuff down. Measure the OD, and put a suitable amount in a
seperate sterile tube. To this add 3 volumes of alcohol, to sterilize the
solution. Put the tube in the hood for half an hour, the liquid should
evaporate (you don't want alcohol on your cells). Then add water and
transfect as usual. 
Qiagen DNA is great for transfections, never had any problem with it,
although I did have a similar white precipitate on occasion.

Kind regards, 

Clemens Suter-Crazzolara, PhD
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