nuclei prep

Jodi Swidzinski jodi.swidzinski at plant-sciences.oxford.ac.uk
Sat Jan 20 04:59:07 EST 2001


No problem.  In fact, we published a short paper on this last year

Folta, K.M. and L.S. Kaufman. (2000) Preparation of transcriptionally
active nuclei from
etilolated Arabidopsis thaliana.  Plant Cell Reports 19:504-10

I used percoll gradients to generate clean nuclei from very small amounts
of etiolated tissue.
The nuceli could be used in the generation of nuclear extracts as well as
for nuclear run-ons to
measure transcription rate.  I do not think it would be difficult to
generate nuclei from tissue culture materials.

The protocol is simple, based on the old pea methods, and gives a
reasonable yield from a small
amount of tissue-- if you have 2g of leaves you will have no problem.

There are a couple of errors in the paper-- our "micros" ended up as
"millis" due to an error in electronic
translation of the text, but I think you can intuitively figure out which
ones they are.

Good luck!

Kevin Folta

""Jodi Swidzinski"" <jodi.swidzinski at plant-sciences.oxford.ac.uk> wrote in
message news:93v50l$nq5$1 at news.ox.ac.uk...
>  Dear All,
>
>  Does anyone have a good protocol for isolating intact, very clean, and
very
>  pure nuclei from Arabidopsis? Cell cultures would be preferable, but any
>  information would help.
>
>  Thanks very much,
>
>  Jodi
>
>
>
>  ---
>
>



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