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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 25mM 4-morpholinepropanesulfonic acid (MOPS), pH 7.0 as a binding buffer, and 25mM MOPS, 1.5M NaCl, 0.1mM 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.

Cryphonectria parasitica; hypovirus; dsRNA; monolith chromatography; detection; purification

3rd Summer School on Monolith Technology for Biochromatography, Bioconversion and Solid-Phase Synthesis