[Arabidopsis] RNA extraction - Arabidopsis lyrata

ulla.kemi from oulu.fi via arab-gen%40net.bio.net (by ulla.kemi from oulu.fi)
Thu Jun 21 03:39:14 EST 2007


I'm studying gene expression in Arabidopsi lyrata. I have some problems
with RNA extraction. I use RNeasy Plant Mini Kit (Qiagen) for extractions.
My leaf samples are stored in RNAlater liquid (Ambion) at -80 degrees. I
do the sample disruption with TissueLyzer (Qiagen): I put the sample,
buffer and 2 stainless steal beads in 2 ml Eppendorf tube and disrupt the
sample by shaking with TissueLyser. For young and fresh leaf samples this
works well: I get good yield of pure RNA. But older and RNAlater stored
samples are causing me problems: the RNA yield is low and it also seems
that there is some impurities in the samples (A260/A230 is really low:
between 0,2 and 1). Does anyone has an idea how should I modify the
extraction protocol to get better quality RNA? And do you have any
experience of working with RNAlater liquid? Could that cause my problems?

Ulla Kemi

Ulla Kemi
University of Oulu
Department of Biology
Plant Genetics Group

email: ulla.kemi from oulu.fi
tel: +358-8-5531795

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