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BMV Propagation, Extraction and Purification Using Chromatographic Methods.

Bio-protocol (2018-07-20)
Aleksander Strugała, Paulina Bierwagen, Jakub Dalibor Rybka, Michał Giersig, Marek Figlerowicz, Anna Urbanowicz
ZUSAMMENFASSUNG

Brome mosaic virus (BMV) is a well-known plant virus representing single-stranded RNA (ssRNA) positive-sense viruses. It has been widely used as a model in multiple studies concerning plant virus biology, epidemiology and the application of viral capsids in nanotechnology. Herein, we describe a method for BMV purification based on ion-exchange and size-exclusion chromatography. The presented method is of similar efficiency to previously described protocols relying on differential centrifugation and can easily be scaled up. The resulting BMV capsids are stable and monodisperse and can be used for further applications.

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Trizma® Base, Primary Standard and Buffer, ≥99.9% (titration), crystalline
Sigma-Aldrich
Ammoniumpersulfat, Molecular Biology, suitable for electrophoresis, ≥98%
Sigma-Aldrich
Natriumphosphat, BioReagent, Molecular Biology, anhydrous, ≥98%
Sigma-Aldrich
Natriumacetat, anhydrous, Molecular Biology, ≥99%
Sigma-Aldrich
Magnesiumchlorid Hexahydrat, BioXtra, ≥99.0%
Sigma-Aldrich
Natriumphosphat, Molecular Biology, ≥98.5% (titration)