Isolation of RNA from wood

[CK Wen]

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
>following aspects:
>
> 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
>        material?
>
> Thanks for your help.
>
>                                Eric
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
approach, IRL)
1.a. You may chop the twig into liquid nitrogen.

1b. Either polytron or grinding could break the tissues.

Q:
 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
low.

regards,

Chi-Kuang