DNA elution from agarose-gel

MICHAEL L SULLIVAN mlsulliv at facstaff.wisc.edu
Fri May 13 09:00:34 EST 2005


I had good luck with QIAEX II to recover large fragments like this (I 
was size selecting a partial digestion for library construction, 8-20 
kb fragments). Make sure to follow the special instructions for large 
fragments. I think I also did a second elution of the resin. I'd 
estimate my recovery was on the order of 30-50%, which I think is more 
than acceptable for such large fragments.

<ole

----- Original Message -----
From: Dirk Müller <Kraweel at web.de>
Date: Friday, May 13, 2005 2:48 am
Subject: DNA elution from agarose-gel

> We have a severe problem with eluting restircted DNA (10-15kb) from
> agarose gel. We have already testet QiaexII, Genomic purification kit
> from Promega, Gel elution colums from Microcon, and electroelution 
but
> don't get more than 1-10 % of the initial DNA which is not enough for
> downstream methods. Using electroelution we can elute the complete 
DNA
> into the membrane(DEAE or hybond N/N+) but we can not elute the DNA
> from the membrane in the elution buffer (15 min at 65°C in 2 M NaCl,
> 10 mM Tris/HCl pH 7 (pH 7-8), 1 mM EDTA). I would welcome any hints
> from this news grout that  will will help with this problem.
> thank you, Dirk...
>
---



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