harvesting RNA from Silique tissue

Dave Yoder yoderda at pilot.msu.edu
Fri Dec 13 10:34:57 EST 2002


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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