DNA isolation from old leaves
chenx071 at umn.edu
Mon Apr 18 10:34:13 EST 2005
Try the conventional CTAB method with higher concentration of NaCl. 1.5M
NaCl is suggested for the first time.
----- Original Message -----
From: "Dr. Klaus Eimert" <eimert at web.de>
To: <plantbio at hgmp.mrc.ac.uk>
Sent: Monday, April 18, 2005 3:18 AM
Subject: Re: DNA isolation from old leaves
> oaktreesrock at yahoo.com (some body) wrote in
> news:20050415152557.67999.qmail at web60623.mail.yahoo.com:
>> Hi everybody,
>> I am having trouble isolating DNA from oak leaves that
>> went from the tree into the -80C freezer about one
>> year ago. Nothing else has been done with them. I am
>> using the Qiagen Plant DNA kit so far, but PCR works
>> only occasionally. Does somebody have a DNA isolation
>> protocol which might give a better chance of getting
>> PCR products?
>> thanks a lot for your help,
>> Do you Yahoo!?
>> Yahoo! Small Business - Try our new resources site!
> Hi Manuel,
> Qiagen states in their handbook that oak leaves result in a very viscous
> lysate with huge amounts of precipitates which may give sheared DNA if
> directly loaded onto the shredder columns. They recommend to introduce an
> additional centrifugation step (handbook page 13). Did you try that?
> So far, we didn't have much luck using commercial kits for grapevine
> leaves (rich in polysaccharides and phenolic compounds - same as oak?).
> Just right now I am testing another bunch of products for that purpose.
> But so far it was always "back to the home brew" ...
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