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

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 ext...

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Veröffentlicht in:Journal of virological methods Jg. 152; H. 1; S. 32 - 37
Hauptverfasser: Balijja, Alitukiriza, Kvarnheden, Anders, Turchetti, Tullio
Format: Journal Article
Sprache:Englisch
Veröffentlicht: London Elsevier B.V 01.09.2008
Amsterdam Elsevier
New York, NY
Schlagworte:
Biological and medical sciences
DNA, Fungal - isolation & purification
DNA, Plant - isolation & purification
dsRNA extraction
dsRNA virus
Fundamental and applied biological sciences. Psychology
Fungi - virology
Microbiology
Nicotiana - virology
RI-dsRNA
RNA Viruses - genetics
RNA, Double-Stranded - isolation & purification
RNA, Viral - isolation & purification
ssRNA virus replicative-intermediate
Techniques used in virology
Virology
dsRNA virus
RI-dsRNA
dsRNA extraction
ssRNA virus replicative-intermediate
Double stranded RNA
Chloroform
Microbiology
Phenols
Method
Virology
ISSN:0166-0934, 1879-0984
Online-Zugang:Volltext
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Zusammenfassung: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 β-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.
Bibliographie:ObjectType-Article-1
SourceType-Scholarly Journals-1
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ISSN:0166-0934
1879-0984
DOI:10.1016/j.jviromet.2008.06.001