DNA isolation from old leaves

Dr. Klaus Eimert eimert at web.de
Mon Apr 18 03:18:32 EST 2005


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,
> 
> Manuel
> 
> 
>           
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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" ...

Cheers,

Klaus 



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