Purification of plant viral and satellite double-stranded RNAs on DEAE monoliths.
Replicative double-stranded RNA (dsRNA) is useful in preliminary identification of Cucumber mosaic virus and its satellite RNA (satRNA). This plant pathogen complex yields sufficient quantity of the replicative RNA form that can be isolated by chromatography on chemically unmodified graded cellulose powder (CF-11). In this work, much faster and more efficient procedure using DEAE monoliths was developed in which dsRNA was separated from other species in total nucleic acids extract originating from the infected plant tissue. The developed chromatographic method revealed the pathogens' presence in only 15 min, avoiding nucleic acid precipitation and electrophoretic analysis.
Krajacić
Mladen
M
University of Zagreb, Faculty of Science, Department of Biology, Rooseveltov trg 6, 10000 Zagreb, Croatia. mladen@botanic.hr
Ivancic-Jelecki
Jelena
J
Forcic
Dubravko
D
Vrdoljak
Anto
A
Skorić
Dijana
D
D002853:Chromatography, Liquid / Q000295:instrumentation / Q000379:methods*; D017799:Cucumovirus / Q000235:genetics*; D004983:Ethanolamines / Q000737:chemistry*; D008958:Models, Molecular; D010942:Plant Viruses / Q000235:genetics; D012330:RNA, Double-Stranded / Q000737:chemistry / Q000302:isolation & purification*; D019244:RNA, Satellite / Q000737:chemistry / Q000302:isolation & purification*; D012367:RNA, Viral / Q000737:chemistry / Q000302:isolation & purification; D015203:Reproducibility of Results
D004983:Ethanolamines; D012330:RNA, Double-Stranded; D019244:RNA, Satellite; D012367:RNA, Viral; C007369:2-diethylaminoethanol