Plant DNA extraction

Barbara E. Liedl bl14 at cornell.edu
Tue Apr 11 08:36:06 EST 1995


In article <cga3-100495105006 at 132.236.3.7> smb14 at cornell.edu writes:
>From: smb14 at cornell.edu
>Subject: Re:  Plant DNA extraction
>Date: Mon, 10 Apr 1995 10:50:06 +0000


>As a quick response to the idea of freeze-drying plant material... We have
>found in our lab that freeze-drying reduces the quality and quantity of DNA
>extracted from the material.  We use fresh tissue most of the time, but
>freeze the tissue at -70C for storage if needed.  It may also depend on the
>tissue.  Brassica is stable and freeze-dries okay, but Malus does not.  We
>beilieve that this is due to greater amount of phenolic compounds in Malus.
> I would suggest doing comparison extractions using fresh, frozen and
>freeze-dried tissue.  

AGREED!  We tried using a protocol from Plant Molecular Biology Reporter by 
Tai and Tanksley (can't remember the year) for tomato.  When we did small runs 
(twelve or less) of the technique it ran okay, not great.  But when we scaled 
up (50+), we ran in to problems.  So, really test the system, we thought we 
had really checked it out, by not only checking the recovery of DNA, but also 
to see if it worked equally well in RFLP & RAPD procedures before switching 
procedures, but were wrong.

We believe the problem was also with the phenolics compounds.  Now we do 
either do fresh or tissue stored at -80.  

Barbara
bl14 at cornell.edu



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