RNA isolation

Dr. Hiranya S. Roychowdhury hroychow at nmsu.edu
Wed Aug 25 12:07:56 EST 2004


Can you grow the fungus in liquid culture?  Usually, for any kind of extraction 
(protein/DNA/RNA), liquid culture (as is done with most ascomycetes in VMM) is 
used.

Alternatively, wash the mycelial/hyphal mass from the solid medium with some 
kind of a buffer or RNase-free water (TE should do) by slow speed (3000rpm) 
centrifugation.  Also, include some sort of RNase inhibitor in your grinding 
buffer and grind the mycelium frozen (LiqN2).  

Transfer ground mycelial powder quickly to the extraction buffer/Phenol mixture 
and proceed with the extraction.  If you are using a commercial kit for it, 
follow their instructions.  Speed often helps. 



Quoting Anja Spielvogel <a.spielvogel at lb.tu-berlin.de>:

> Hi,
> 
> I try to isolate RNA from a filamentous fungi grown on agar plates. When I
> grow the fungus on Cellophan it changes growth physiology and expression
> pattern. I´d like to avoid growing on Cellophan, but RNA isolation becomes
> complicated, because RNA is degraded when I scrape the fungus or try RNA
> isolation with whole agar pieces.
> Any suggestions?
> 
> Thanks in advance
> 
> Anja
> 
> 


-- 
Hiranya S. Roychowdhury, Ph.D.
Coll. Asst. Professor,
Molecular Biology,
Dept. of Chemistry & Biochemistry
Rm# 336, Chemistry Bldg.; MSC 3MLS
New Mexico State University
Las Cruces, NM 88003
---



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