Growth Recovery Assay and FACS-based Population Sorting Following Territorial Exclusion in Proteus mirabilis
Year: 2020
Type: article
Abstract: Ids system, are non-lethal and do not cause obvious physiological changes in target cells. Previously, experimental techniques to probe cells experiencing non-lethal kin recognition have been limited. Here we describe a technique to reliably isolate cells deemed self and non-self through Ids self-recognition for downstream phenotypic analysis. Liquid cultures of fluorescently labeled self-recognition mutants are mixed together and inoculated on swarm-permissive agar. Mixed swarms are harvested, and each strain is isolated through fluorescence-activated cell sorting (FACS). The growth rate of each strain is measured on a plate reader. This protocol is adaptable for other bacterial species. We describe briefly how sorted particles can be used for other analyses such as RNA-Seq library preparation.
Source: BIO-PROTOCOL
Authors: Murray Tipping, Karine Gibbs
Institution: Harvard University
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: green
OpenAlex ID: W3010471590