Need help for emoving glycogen from DNA solution
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).
c/o bhandary at wccf.mit.edu
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