[Arabidopsis] RE: agar-grown roots. Arab-gen digest, vol 12, issue 2

Lorvellec, Maelle Maelle.Lorvellec at wur.nl
Fri Apr 7 05:20:31 EST 2006

Message: 1
Date: Fri, 31 Mar 2006 15:42:15 -0500
From: "Redman, Julia C." <jredman at tigr.ORG>
Subject: [Arabidopsis] Agar-grown roots for RNA
To: <arab-gen at magpie.bio.indiana.edu>
        <7F0AA25C9B1706448E8B73DFDD109B3102453795 at EXCHANGE.TIGR.ORG>
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Hello everyone,

We're looking to do some expression profiling using root tissue grown in
MS Agar and I was wondering about the logistics of plucking roots from
agar for RNA extraction.  What is the best way to go about this without
breaking them?

Any advice will be appreciated!

Julia Redman

The Institute for Genomic Research
Dear Julia,

For isolating RNA, I sow the seeds in 1 ligne on 1 plate of 1/2 MS  and I grow them vertically so
the roots grow on top of the agar. Then with a knife I cut all the roots in one time at the
same level and harvest them for RNA. 
You can also still grow them horizontaly in 1/2 MS plate and pull them very gently but then they
should not grow for more than a week and you need to cut each root individually.
1/2 MS = 2.2g/L Murashige and Skoog 10 +vitamins, 10 g/L sucrose, pH 5.8 with KOH, 0.8% Diashin agarose.

Hope it helps. If you have more questions about RNA isolation please let me know.


Laboratory of Molecular Biology.Plant Sciences.Wageningen University.Netherlands.



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