[en] Outer membrane vesicles (OMVs) are released by commensal as well as pathogenic Gram-negative bacteria. These vesicles contain numerous bacterial components, such as proteins, peptidoglycans, lipopolysaccharides, DNA, and RNA. To examine if OMV-associated RNA molecules are bacterial degradation products and/or are functionally active, it is necessary to extract RNA from pure OMVs for subsequent analysis. Therefore, we describe here an isolation method of ultrapure OMVs and the subsequent extraction of RNA and basic steps of RNA-Seq analysis. Bacterial culture, extracellular supernatant concentration, OMV purification, and the subsequent RNA extraction out of OMVs are described. Specific pitfalls within the protocol and RNA contamination sources are highlighted.
Centre de recherche :
- Luxembourg Centre for Systems Biomedicine (LCSB): Eco-Systems Biology (Wilmes Group) - Luxembourg Centre for Systems Biomedicine (LCSB): Bioinformatics Core (R. Schneider Group) ULHPC - University of Luxembourg: High Performance Computing
Disciplines :
Microbiologie
Auteur, co-auteur :
HABIER, Janine ; University of Luxembourg > Luxembourg Centre for Systems Biomedicine (LCSB)
MAY, Patrick ; University of Luxembourg > Luxembourg Centre for Systems Biomedicine (LCSB)
Heintz-Buschart, Anna
Ghosal, Anubrata
WIENECKE-BALDACCHINO, Anke ; University of Luxembourg > Faculty of Science, Technology and Communication (FSTC) > Life Science Research Unit
Nolte-'t Hoen, Esther N.M
WILMES, Paul ; University of Luxembourg > Luxembourg Centre for Systems Biomedicine (LCSB)
FRITZ, Joëlle ; University of Luxembourg > Luxembourg Centre for Systems Biomedicine (LCSB)
Co-auteurs externes :
yes
Langue du document :
Anglais
Titre :
Extraction and Analysis of RNA Isolated from Pure Bacteria-Derived Outer Membrane Vesicles
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