DNA extraction of 140 kb piece from agarose

Michael Szardenings msz at bio.embnet.se
Fri Nov 10 06:40:16 EST 1995


In article <DHJDrs.4n9 at freenet.carleton.ca>, bm335 at FreeNet.Carleton.CA
(Christina Chan) wrote:

> I have been having difficulty in isolating a 140 kb piece of DNA
> from a TBE agarose (1% SeaKem GTG) plug.  The plug contains
> ~500 ng of DNA.  I have tried the Gene Clean II kit but was un-
> successful.  
> 
> Any suggestions?
If it is 140 bp, then you will really have problems with most silica based
kits. Use a kit designed for PCR fragment purification. I used 'Jetpure'
from Genomed and they guarantee, that you loose 100% of all primers etc
below 50 bp. Thus the yield in your case might not even be the best with
such kits, but it worked well for me in most cases.
Besides, you might be more successful with electroelution, its the ideal size.
Good luck
      Michael

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