@inbook{b057763726374533a832d9400aeaf58e,
title = "Extraction and analysis of RNA isolated from pure bacteria-derived outer membrane vesicles",
abstract = "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.",
keywords = "Analysis, Bacteria, Density gradient, Extraction, Gram-negative, Outer membrane vesicle (OMV), RNA, Sequencing, Ultracentrifugation, Ultrafiltration",
author = "Janine Habier and Patrick May and Anna Heintz-Buschart and Anubrata Ghosal and Wienecke-Baldacchino, {Anke K.} and {Nolte-{\textquoteleft}t Hoen}, {Esther N.M.} and Paul Wilmes and Fritz, {Jo{\"e}lle V.}",
note = "Publisher Copyright: {\textcopyright} 2018, Springer Science+Business Media, LLC.",
year = "2018",
doi = "10.1007/978-1-4939-7634-8_13",
language = "English",
series = "Methods in Molecular Biology",
publisher = "Humana Press Inc.",
pages = "213--230",
booktitle = "Methods in Molecular Biology",
}