Separation of plant viral satellite double-stranded RNA using high-performance liquid chromatography.
High-performance liquid chromatography was developed for further separation of double-stranded (ds) RNAs obtained by CF-11 cellulose chromatography from plants infected with satellite associated cucumber mosaic virus. Fractions separated by monolithic polymer column, especially applicable for nucleic acid analyses, were identified electrophoretically and confirmed with a polymerase chain reaction test. Once standardized, the method has revealed clear evidence of satellite presence without precipitation and electrophoresis. According to demonstrated sensitivity, its application in the preliminary diagnostics of field samples is also predictable. Principally, it can be used as a powerful preparative approach resulting in highly pure satellite dsRNA for further analyses.
Likić
Sasa
S
University of Zagreb, Faculty of Science, Department of Biology, Rooseveltov trg 6, 10000 Zagreb, Croatia.
Rusak
Gordana
G
Krajacić
Mladen
M
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D017799:Cucumovirus / Q000235:genetics; D010935:Plant Diseases / Q000821:virology; D012330:RNA, Double-Stranded / Q000302:isolation & purification*; D019244:RNA, Satellite / Q000302:isolation & purification*
D012330:RNA, Double-Stranded; D019244:RNA, Satellite