[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


Hi,

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?



Thanks,
Ulla Kemi



-------------
Ulla Kemi
University of Oulu
Department of Biology
Plant Genetics Group
Finland

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



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