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, 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.
Perica
Mirna Curković
MC
University of Zagreb, Faculty of Science, Department of Biology, Rooseveltov trg 6, 10000 Zagreb, Croatia.
Sola
Ivana
I
Urbas
Lidija
L
Smrekar
Franci
F
Krajacić
Mladen
M
D001203:Ascomycota / Q000821:virology*; D002852:Chromatography, Ion Exchange / Q000379:methods*; D012328:RNA Viruses / Q000737:chemistry / Q000302:isolation & purification*; D012330:RNA, Double-Stranded / Q000302:isolation & purification*
D012330:RNA, Double-Stranded