Subject: Re: Isolation of RNA from Rhizoctonia solani
From: [email-masked] (Seanna Annis)
Date: 8 Jul 1994 19:39:36 GMT
Newsgroups: bionet.mycology

Ethan Strauss ([email-masked]) wrote:
: I need to isolate mRNA from R. solani. Ideally I would like
: to get rid of double-stranded RNA's, but I can live with them
: if I need to. Does anyone have any refrences, protocols, advice ect?
: Thanks for your help.
: Ethan Strauss                              [email-masked]
: Dept. Plant Biology & Pathology            74 N. Main St. Apt#1
: University of Maine at Orono 04469         Orono ME, 04473
: 207-581-2994                               207-866-5935

I have used Trizol reagent, from Gibco BRL to isolate total RNA from 
Leptosphaeria maculans mycelium with good results.  The method is fast, 
around 2 hours, and gives lots of clean RNA (only small amount of 
DNA).  I have not isolated mRNA from this RNA but considering the high 
yields I think it would work well.  I got between 30 to 90 ug of RNA for
 each 100 mg of fresh mycelium I used.  This RNA worked well for first
 strand cDNA synthesis and RT-PCR afterwards.  As you can tell I really 
liked this reagent, I thought it would take me days to get RNA using the 
CsCl method.     Seanna Annis, [email-masked]