A non-phenol-chloroform extraction of double-stranded RNA from plant and fungal tissues

Abstract

Double-stranded RNA (dsRNA) molecules of viruses are found in nature at a very high frequency. Their detection in plants and fungi has been carried out with difficulty due to the complicated dsRNA extraction techniques used commonly which includes phenol-chloroform extractions. In this study, an extraction method for isolation of dsRNA is described that is free of phenol and chloroform. A lysis buffer, containing P-mercaptoethanol and polyvinylpolypyrrolidone (PVPP-40), was added to homogenised tissues and the subsequent supernatant was filtered through a cellulose CF-11 mini-column. DsRNA molecules were separated based on the differing affinity of nucleic acids for the cellulose CF-11 resin in 20% ethanol buffer. This easy, rapid and cheap technique has been successfully tested on fungi and plants containing different dsRNA virus molecules, indicating the possibility of a wide use of the method. (C) 2008 Elsevier B.V. All rights reserved.

Keywords

dsRNA virus; dsRNA extraction; ssRNA virus replicative-intermediate; RI-dsRNA

Published in

Journal of Virological Methods
2008, Volume: 152, number: 1-2, pages: 32-37
Publisher: ELSEVIER SCIENCE BV

SLU Authors

• Kvarnheden, Anders

  • Department of Plant Biology, Swedish University of Agricultural Sciences
    

UKÄ Subject classification

Agricultural Science


Publication identifier

DOI: https://doi.org/10.1016/j.jviromet.2008.06.001

Permanent link to this page (URI)

https://res.slu.se/id/publ/18534