glycogen carrier for nucleic acid ppt

bugg at mbcf.stjude.org bugg at mbcf.stjude.org
Fri Dec 20 10:33:24 EST 1991


In article <1991Dec18.232950.6402 at usenet.ins.cwru.edu>, ccy at po.CWRU.Edu (Cheung C. Yue) writes:
> 
> I am interested in using glycogen as a carrier for nucleic acids
> ppt.  Tracy (Prep Biochem 11:251, '81) described purification of
> glycogen for such use.  Steps include micrococcal nuclease, proteinase
> K, then extensive phenol extraction.  Another paper I've seen 
> said they just took glycogen from Sigma, dissolved it in water and
> use.  Does anyone have any experience with this problem?  Are there
> problems with the second approach?
> 
> ccy at po.cwru.edu
>   
> -- 
I use this method for precipitation with glycogen and it is very effective for
recovery of very small DNA (such as PCR products <200 bp).  I use Sigma
glycogen type II cat # G-8751 and make a 10 mg/ml stock in sterile water.  I
don't do any type of extraction, but the author you mentioned may have been
trying to prepare it for a different purpose.  For example, if you were
planning to use glycogen to precipitate DNA from forensic samples and then do
PCR, you would probably want to get rid of any DNA that might be present in it
(thus, the nuclease that Tracy added).  I first got the idea of using glycogen
from a paper in which DNA was precipitated from blood stains and then used for
PCR.  Anyway, I add 1/50 volume of my glycogen stock + 2 volumes of -20C EtOH
and precipitate on ice for 10 min.  Microfuging for 15 min gives a large white
pellet, which I dry and resuspend to load on a agarose gel.  Also, glycogen
does not seem to interfere with T4 DNA ligase or any of the restriction enzymes
I use.  Good luck!
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Barbara Bugg, M.S.  bugg at mbcf.stjude.org       "I refuse to have a battle of
St. Jude Children's Research Hospital          wits with an unarmed person."
Memphis,  TN  USA
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