Purifying DNA from gel slices

Chung Jui Tsai chtsai at MTU.EDU
Sat Nov 9 20:49:45 EST 1996


At 11:51 PM 11/9/96 GMT, you wrote:
>Alison Reavley <uzdb0017 at sable.ox.ac.uk> sez:
>
>
>: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.
>
>
>This may or may not be helpful -- certainly GeneClean is out of the
>question because of the temperature criterion -- but there was an
>article in BioTechniques several years ago (I'll try to find it) where
>the agarose chunk is placed in a syringe with some chaotropic agent
>and pushed through a standard filter...then I think you just have to
>precipitate it. Anybody know it? I'll look for the article. ;)
>
>cp
>                  
>   _o    Cogito ergo sum is nonsense; I think _in spite_ 
> _ \,_   of being, and I _am_ in spite of thinking.
>(_)/(_)  
>
>

The references are :
Li and Ownby 1993 BioTechniques 15 (6): 976-978.
Lu et al. 1994 BioTechniques 16(3):400-402.
There was another article in BioTechniques recently (couple months ago) in
which a filter-tip is used (placed in a microcentrifuge tube) instead.

C.J. Tsai




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