Need help for emoving glycogen from DNA solution

Uttam Rajbhandary bhandary at wccf.mit.edu
Wed Dec 7 18:15:00 EST 1994


In article <3bhs48$a6h at obelix.cica.es>, .bb1roara at lucano.uco.es (Antonio Rodriguez-Ariza) writes...
>I am having problems in the purification of total DNA from fish (Sparus
>aurata, gilthead seabream) liver. Using two distinct protocols
>(phenol/chloroform or benzylchloride extraction), I always obtain a DNA
>solution but with a high amount of GLYCOGEN. How can I remove glycogen
>from this DNA solution?

After redissolving your ethanol precipitate, you may want to try adjusting the 
sample to 0.3 M NaAc (neutral), and then spining at 10-20000 x g for 20 
minutes.  The glycogen should pellet.  I run into this problem depending upon 
the strain with making single-strand templates from phagemid vectors.  Certain 
strains gives DNA that seems to be contaminated with some sort of high 
molecular carbohydrate-like material.  I don't think it's polyethylene glycol, 
since it appears to be strain-related (in my hands, XS127 is a great 
offender).  The ethanol pellet is white, and larger than the OD that one gets. 
 Also, there is always an interphase with phenol-chloroform extraction.  The 
material pelleted as I describe above has no A260.
I can't remember where I saw the original method for removing the glycogen, 
which is the connection I thought of to try this in the first place).

Harold Drabkin
c/o bhandary at wccf.mit.edu




More information about the Methods mailing list