Purification of G1 daughter cells from different Saccharomycetes species through an optimized centrifugal elutriation procedure.

Yeast

Institut Pasteur, Group Spatial Regulation of Genomes, Department of Genomes and Genetics, F-75015, Paris, France; CNRS, UMR3525, F-75015, Paris, France.

Published: May 2014

Centrifugal elutriation discriminates cells according to their sedimentation coefficients, generating homogeneous samples well suited for genomic comparative approaches. It can, for instance, isolate G1 daughter cells from a Saccharomyces cerevisiae unsynchronized population, alleviating ageing and cell-cycle biases when conducting genome-wide/single-cell studies. The present report describes a straightforward and robust procedure to determine whether a cell population of virtually any yeast species can be efficiently elutriated, while offering solutions to optimize success. This approach was used to characterize elutriation parameters and S-phase progression of four yeast species (S. cerevisiae, Candida glabrata, Lachancea kluyveri and Pichia sorbitophila) and could theoretically be applied to any culture of single, individual cells.

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http://dx.doi.org/10.1002/yea.3005DOI Listing

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