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harvesting RNA from Silique tissue

Dr. James Campanella campanellj at mail.montclair.edu
Sat Dec 14 07:08:19 EST 2002


Julia,

David is correct. We have been using Qiagen's RNeasy to extract RNA from a
whole variety of Arabidopsis tissues for the last few months-- roots,
shoots, basal
leaves, apical leaves, and even hypocotyls. We have gotten very consistent,
very clean
results with the kit. It is worth the money. I would also invest in RNA
Secure which
an anti-RNase reagent from Ambion. We do no extractions w/o RNA Secure
included.
It's wonderful because it is formulated to not interfere with other
biochemical
reactions and can be left in as insurance for procedures like extraction or
even
RT-PCR.

Good luck,

Jim


At 03:34 PM 12/13/2002 +0000, Dave Yoder wrote:
>I have been using Qiagen's Plant RNeasy columns to extract total RNA from
8-10
>day old plate grown Arabidopsis shoots.  I have not specifically tried to
>isolate total RNA from silique tissue.  Hovever, in a Northern blot
comparison
>between total RNA isolated using RNeasy columns and TRIzol method, the RNeasy
>method gives me consistant RNA yeilds of 2-3.5 ug/ul with no degradation.
The
>total RNA isolated using TRIzol was almost completely degraded.  I
>would suggest
>that you try the Qiagen Plant RNeasy kit as it is what other labs here at MSU
>are using to prepare total RNA for microarray analysis.
>
>--
>Dr. David Yoder
>Department of Plant Biology
>166 Plant Biology Building
>Michigan State University
>East Lansing, MI 48824-1312
>
>"Redman, Julia" wrote:
>
>>   <x-charset iso-8859-1>Good afternoon, everyone.
>>
>>   Recently I made a second attempt at harvesting total RNA from silique
tissue
>>   using TRIzol reagent and Invitrogen's protocol, followed by a
polysaccharide
>>   precipitation.  Analysis indicates the only thing I was able to recover
was
>>   tRNA.  Can anyone share tips or a better protocol for extracting total RNA
>>   from samples such as siliques?
>>
>>   I appreciate!
>>
>>   Julia
>>
>>   ____________________________________________
>>   Julia Redman
>>   The Institute for Genomic Research
>>   9712 Medical Center Dr.
>>   Rockville, MD 20850
>>
>>   To have me paged at TIGR:  301-838-0200
>>
>>   ---
>>
>>   </x-charset>
>
>
>
>
>
>---
>

---



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