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 ![]() | |
May, Patrick ![]() | |
Heintz-Buschart, Anna [] | |
Ghosal, Anubrata [] | |
Wienecke-Baldacchino, Anke ![]() | |
Nolte-'t Hoen, Esther N.M [] | |
Wilmes, Paul ![]() | |
Fritz, Joëlle ![]() | |
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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