Isolation of RNA from wood
cwen at aux.btny.purdue.edu
Mon Feb 5 09:21:00 EST 1996
hamburg at sage.cc.purdue.edu (Eric Sarnighausen) wrote:
>I am interested in the isolation of functional RNA from the xylem of red
>osier dogwood (Cornus sericea L.) twigs. Only the youngest twigs of
>which the maximum diameter is 0.3 inches will be used.
>I would be extremely interested to obtain information regarding the
> I.) Optimizing the total amount of extractable RNA (cell breakage)
> a.) What is the best way to pretreat (cut, rasp) the wood before it
> is frozen and stored?
> b.) How is the mechanical cell breakage prior to/at the beginning of
> the extraction process to be accomplished most efficiently?
> c.) Is it possible to isolate functional RNA (in acceptable amounts)
> from material that has been freeze-dried (and is therefore easy
> to grind)?
> II.) Optimizing the quality of the isolated RNA
> What potential problems regarding the extraction process itself
> have to be taken into account when RNA is extracted from woody
> Thanks for your help.
1.c. Many people use frozen-dried tissues for nucleic acids preparation and the nucleid acids are preserved much better because the solvent, water, has been removed. For lyopholized tissues, they can be stored at
-70C for a very long period. ( ref: Plant Molecular Biology, a practical
1.a. You may chop the twig into liquid nitrogen.
1b. Either polytron or grinding could break the tissues.
Is the xylem dead tissue? How could you isolate functional RNA or RNA
related to the function of xylem?
II. The potential problem could be low yield because the tissue contains a huge amount of dead cells and the ratio of live cells to dead cells is
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