Naslov
Purification of Plant Viral and Satellite dsRNAs on CIM&reg ; ; : DEAE Monoliths

Autori
Krajačić, Mladen ; Ivančić-Jelečki, Jelena ; Forčić, Dubravko ; Vrdoljak, Anto ; Škorić, Dijana

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Izvornik
2006 Summer School Abstract Book / Aleš Štrancar - Ljubljana : BIA Separations, 2006, 15-16

Skup
Monolith Summer School 2006

Mjesto i datum
Portorož, Slovenija, 28.05.2006. - 02.06.2006

Vrsta sudjelovanja
Pozvano predavanje

Vrsta recenzije
Međunarodna recenzija

Ključne riječi
convective interaction media; cucumber mosaic virus (CMV); CMV satRNA; preliminary diagnosis

Sažetak
Most plant viruses possess single-stranded (+) RNA as their genome. Its replication generates double-stranded form (dsRNA), which is useful in diagnosis and can be isolated by CF-11 cellulose chromatography. This approach is often used in preliminary identification of cucumber mosaic virus and associated satellite RNA, sometimes a very destructive pathogen combination distributed worldwide in tomato. In this work, much faster and more efficient dsRNA isolation procedure using CIM&reg ; ; ; DEAE monolithic columns was developed. By optimizing the binding and elution conditions, satisfactory chromatographic separation and purification of pathogens’ replicative forms was achieved including separation of satellite from viral dsRNAs. Subsequent electrophoretic analyses, PCR and RNase-treatment assays confirmed the contents of the fractions. Once standardized, this procedure results in chromatograms that confirm viral and satellite presence, avoiding dsRNA precipitation and electrophoresis, while fractions can be used for further analyses of the pathogen complex components.

Izvorni jezik
Engleski

Znanstvena područja
Biologija

Napomena
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