Isolation of intact RNA from cytometrically sorted Saccharomyces cerevisiae for the analysis of intrapopulation diversity of gene expression

authored by

Jeannette Achilles, Frank Stahl, Hauke Harms, Susann Müller

Abstract

Characterizing and understanding the functional heterogeneity in a given population on the cellular and molecular level is a great challenge in microbiology. Each microorganism contributes differently to the overall performance of the community and responds differently to changing microenvironmental conditions. Here, we present a method for isolation of intact RNA out of small subpopulations of live Saccharomyces cerevisiae cells for differential gene expression analysis. The protocol includes fluorescence staining, flow cytometric analysis and sorting of live yeast cells, subsequent isolation of RNA from the resulting subpopulations and finally RNA quantification and integrity check. The isolated RNA can be transcribed into cDNA and successfully used for microarray analysis. This aids in relating molecular regulation processes within subpopulations with the dynamics and functioning of the entire population. The procedure can be accomplished in 2 d.

Organisation(s)

Institute of Technical Chemistry

External Organisation(s)

Helmholtz Centre for Environmental Research (UFZ)

Type

Article

Journal

Nature protocols

Volume

2

Pages

2203-11

No. of pages

9

ISSN

1750-2799

Publication date

2007

Publication status

Published

Peer reviewed

Yes

Sustainable Development Goals

SDG 3 - Good Health and Well-being

Electronic version(s)

https://doi.org/10.1038/nprot.2007.322 (Access: Unknown)