Reference : Extraction and Analysis of RNA Isolated from Pure Bacteria-Derived Outer Membrane Vesicles
Parts of books : Contribution to collective works
Life sciences : Microbiology
Systems Biomedicine
http://hdl.handle.net/10993/35056
Extraction and Analysis of RNA Isolated from Pure Bacteria-Derived Outer Membrane Vesicles
English
Habier, Janine mailto [University of Luxembourg > Luxembourg Centre for Systems Biomedicine (LCSB) > >]
May, Patrick mailto [University of Luxembourg > Luxembourg Centre for Systems Biomedicine (LCSB) > >]
Heintz-Buschart, Anna []
Ghosal, Anubrata []
Wienecke-Baldacchino, Anke mailto [University of Luxembourg > Faculty of Science, Technology and Communication (FSTC) > Life Science Research Unit >]
Nolte-'t Hoen, Esther N.M []
Wilmes, Paul mailto [University of Luxembourg > Luxembourg Centre for Systems Biomedicine (LCSB) > >]
Fritz, Joëlle mailto [University of Luxembourg > Luxembourg Centre for Systems Biomedicine (LCSB) > >]
27-Feb-2018
Methods in Molecular Biology
Bacterial Regulatory RNA
Arluison, Véronique
Valverde, Claudio Valverde
Humana Press
1737
213-230
Yes
978-1-4939-7634-8
New York
USA
[en] Bacteria ; RNA ; Outer membrane vesicle (OMV) ; Gram-negative ; Sequencing ; Extraction ; Ultracentrifugation ; Ultrafiltration ; Density gradient
[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.
Luxembourg Centre for Systems Biomedicine (LCSB): Eco-Systems Biology (Wilmes Group) ; Luxembourg Centre for Systems Biomedicine (LCSB): Bioinformatics Core (R. Schneider Group) ; University of Luxembourg: High Performance Computing - ULHPC
Researchers
http://hdl.handle.net/10993/35056
10.1007/978-1-4939-7634-8_13
https://link.springer.com/protocol/10.1007%2F978-1-4939-7634-8_13
FnR ; FNR8066232 > Joelle Fritz > BEaR > Host-microbes interactions via bacterial extracellular small RNAs > 01/04/2015 > 31/08/2017 > 2014

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