Separation of cells in density gradients with temporary solidification of the gradient: Application to Escherichia coli forms obtained by microwave treatment of pure cultures
: Abashina T., Vainshtein M., Korpela S., Korpela T.
Publisher: Elsevier B.V.
: 2018
Journal of Microbiological Methods
: Journal of Microbiological Methods
: 153
: 45
: 47
: 3
: 0167-7012
: 1872-8359
DOI: https://doi.org/10.1016/j.mimet.2018.09.003
Gelatin added in certain concentration into the density gradient mixtures allows to solidify and melt the gradient reversibly. Such a method was applied to separation of partially disintegrated Escherichia coli VKM B-125 cells in discontinuous sucrose gradient which was optimized to obtain small round “nano-cells” free from other cell components.
Gelatin added to the sucrose solutions had the advantages: i) accurate gradient layers were formed, ii) gelatin did not interfere with the cells and sub-particles during centrifugation, iii) solidified gelatin allowed to cut out the separated cell components, and iv) gelatin allowed to prepare ready-made solidified gradient centrifugation tubes into storage.