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

Sammanfattning

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.

Nyckelord

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

Publicerad i

Journal of Virological Methods
2008, volym: 152, nummer: 1-2, sidor: 32-37
Utgivare: ELSEVIER SCIENCE BV

SLU författare

• Kvarnheden, Anders

  • Institutionen för växtbiologi, Sveriges lantbruksuniversitet
    

UKÄ forskningsämne

Jordbruksvetenskap

Publikationens identifierare

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

Permanent länk till denna sida (URI)

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