Publication Details |
| Category | Text Publication |
| Reference Category | Book chapters |
| DOI | 10.1007/978-1-4939-7634-8_13 |
| Title (Primary) | Extraction and analysis of RNA isolated from pure bacteria-derived outer membrane vesicle |
| Title (Secondary) | Bacterial regulatory RNA : methods and protocols |
| Author | Habier, J.; May, P.; Heintz-Buschart, A.; Ghosal, A.; Wienecke-Baldacchino, A.K.; Nolte-‘t Hoen, E.N.M.; Wilmes, P.; Fritz, J.V. |
| Publisher | Arluison, V.; Valverde, C. |
| Journal | Methods in Molecular Biology |
| Year | 2018 |
| Department | BOOEK; iDiv |
| Volume | 1737 |
| Page From | 213 |
| Page To | 230 |
| Language | englisch |
| Keywords | Bacteria; RNA; Outer membrane vesicle (OMV); Gram-negative; Sequencing; Analysis; Extraction; Ultracentrifugation; Ultrafiltration; Density gradient |
| 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. |
| Persistent UFZ Identifier | https://www.ufz.de/index.php?en=20939&ufzPublicationIdentifier=21384 |
| Habier, J., May, P., Heintz-Buschart, A., Ghosal, A., Wienecke-Baldacchino, A.K., Nolte-‘t Hoen, E.N.M., Wilmes, P., Fritz, J.V. (2018): Extraction and analysis of RNA isolated from pure bacteria-derived outer membrane vesicle In: Arluison, V., Valverde, C. (eds.) Bacterial regulatory RNA : methods and protocols Methods in Molecular Biology 1737 Springer Nature, p. 213 - 230 |
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