While different giant viruses' purification protocols are available, they are not fully described and they use sucrose gradient that does not reach an equilibrium. Here, we report a protocol for the purification of members of the Mimiviridae family virions resulting from Acanthamoeaba castellanii infections. Viruses are harvested after cell lysis and purified through a high density CsCl gradient to optimize the isolation of the virus from the cell debris or other potential contaminants. Due to the large size of the virion capsids, reaching half a micrometer diameter, the quality of the process can be monitored by light microscopy. The resulting purified particles can then be used to perform new infections, DNA extraction, structural studies, sugar composition analyses, sub-compartment characterization or proteomic experiments. Copyright © 2020 The Authors; exclusive licensee Bio-protocol LLC.
Citation
Lionel Bertaux, Audrey Lartigue, Sandra Jeudy. Giant Mimiviridae CsCl Purification Protocol. Bio-protocol. 2020 Nov 20;10(22):e3827
PMID: 33659479
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