Separation of hypoviral double-stranded RNA on monolithic chromatographic supports

A procedure based on BIA Separations CIM DEAE anion-exchange chromatography was developed to separate double-stranded (ds) RNA of hypovirus infecting phytopathogenic fungus Cryphonectria parasitica. Using a linear gradient of 25 mM 4-morpholinepropanesulfonic acid (MOPS), pH 7.0 as a binding buffer,...

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Bibliographic Details
Published in:Journal of Chromatography A 2009-03, Vol.1216 (13), p.2712-2716
Main Authors: Perica, Mirna Ćurković, Šola, Ivana, Urbas, Lidija, Smrekar, Franci, Krajačić, Mladen
Format: Article
Language:English
Subjects:
chromatography
Cryphonectria parasitica
Cryphonectria parasitica hypovirus
Detection
double-stranded RNA
dsRNA
Hypovirus
Monolith chromatography
Purification
separation
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Summary:A procedure based on BIA Separations CIM DEAE anion-exchange chromatography was developed to separate double-stranded (ds) RNA of hypovirus infecting phytopathogenic fungus Cryphonectria parasitica. Using a linear gradient of 25 mM 4-morpholinepropanesulfonic acid (MOPS), pH 7.0 as a binding buffer, and 25 mM MOPS, 1.5 M NaCl, 0.1 mM EDTA, 15% isopropanol (v/v), pH 7.0 as an elution buffer, hypoviral dsRNA was additionally purified from nucleic acid species present in preparations partially purified by standard CF-11 cellulose chromatography. Moreover, crude phenol/chloroform extracts of the fungal tissue were also applied to monolithic supports and CIM DEAE chromatograms revealed clear evidence for hypoviral presence without CF-11 chromatography, nucleic acid precipitation, and electrophoresis.
ISSN:0021-9673
DOI:10.1016/j.chroma.2008.10.115