DNA extractions

Steven Casper casper at scripps.edu
Fri Dec 1 07:22:37 EST 1995


In article <49n9o2$7tv at mserv1.dl.ac.uk>, "Corinna. Dutton"
<Corinna.Dutton at newcastle.ac.uk> wrote:

> I have had problems purifying genomic DNA .........



   Hello There,

   I have forwarded your post to a coworker who has a protocol for DNA
extraction from maize endosperm.  In the meantime I would suggest that you
try an organic extraction of the isolated DNA using an equal volume of 1:1
chloroform:butanol.  This has helped me remove most of the polysaccharides
from my preps.  
   My protocol comes from Current Protocols in Molecular Biology and
consists of  freezing the tissue, grinding, adding extraction buffer
(Tris, EDTA, NaCl, Proteinase K), adding Sarkosyl, spinning out
debris,.........
   In the comentary of the protocol, polysaccharide removal is recommended
by chloroform extraction in the presence of 1% CTAB and 0.7 M NaCl but I
do not include this in my preps so I can not vouch for its effectiveness. 

   If you would like, I could send a detailed protocol.  Just let me know.

   Good Luck, 

   Steve



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