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Incomplete cell disruption of resistant microbes

Biomolecules for OMIC analysis of microbial communities are commonly extracted by bead-beating or ultra-sonication, but both showed varying yields. In addition to that, different disruption pressures are necessary to lyse bacteria and fungi. However, the disruption efficiency and yields comparing be...

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Autores principales: Starke, Robert, Jehmlich, Nico, Alfaro, Trinidad, Dohnalkova, Alice, Capek, Petr, Bell, Sheryl L., Hofmockel, Kirsten S.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6449382/
https://www.ncbi.nlm.nih.gov/pubmed/30948770
http://dx.doi.org/10.1038/s41598-019-42188-9
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author Starke, Robert
Jehmlich, Nico
Alfaro, Trinidad
Dohnalkova, Alice
Capek, Petr
Bell, Sheryl L.
Hofmockel, Kirsten S.
author_facet Starke, Robert
Jehmlich, Nico
Alfaro, Trinidad
Dohnalkova, Alice
Capek, Petr
Bell, Sheryl L.
Hofmockel, Kirsten S.
author_sort Starke, Robert
collection PubMed
description Biomolecules for OMIC analysis of microbial communities are commonly extracted by bead-beating or ultra-sonication, but both showed varying yields. In addition to that, different disruption pressures are necessary to lyse bacteria and fungi. However, the disruption efficiency and yields comparing bead-beating and ultra-sonication of different biological material have not yet been demonstrated. Here, we show that ultra-sonication in a bath transfers three times more energy than bead-beating over 10 min. TEM imaging revealed intact gram-positive bacterial and fungal cells whereas the gram-negative bacterial cells were destroyed beyond recognition after 10 min of ultra-sonication. DNA extraction using 10 min of bead-beating revealed higher yields for fungi but the extraction efficiency was at least three-fold lower considering its larger genome. By our critical viewpoint, we encourage the review of the commonly used extraction techniques as we provide evidence for a potential underrepresentation of resistant microbes, particularly fungi, in ecological studies.
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spelling pubmed-64493822019-04-10 Incomplete cell disruption of resistant microbes Starke, Robert Jehmlich, Nico Alfaro, Trinidad Dohnalkova, Alice Capek, Petr Bell, Sheryl L. Hofmockel, Kirsten S. Sci Rep Article Biomolecules for OMIC analysis of microbial communities are commonly extracted by bead-beating or ultra-sonication, but both showed varying yields. In addition to that, different disruption pressures are necessary to lyse bacteria and fungi. However, the disruption efficiency and yields comparing bead-beating and ultra-sonication of different biological material have not yet been demonstrated. Here, we show that ultra-sonication in a bath transfers three times more energy than bead-beating over 10 min. TEM imaging revealed intact gram-positive bacterial and fungal cells whereas the gram-negative bacterial cells were destroyed beyond recognition after 10 min of ultra-sonication. DNA extraction using 10 min of bead-beating revealed higher yields for fungi but the extraction efficiency was at least three-fold lower considering its larger genome. By our critical viewpoint, we encourage the review of the commonly used extraction techniques as we provide evidence for a potential underrepresentation of resistant microbes, particularly fungi, in ecological studies. Nature Publishing Group UK 2019-04-04 /pmc/articles/PMC6449382/ /pubmed/30948770 http://dx.doi.org/10.1038/s41598-019-42188-9 Text en © The Author(s) 2019 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Starke, Robert
Jehmlich, Nico
Alfaro, Trinidad
Dohnalkova, Alice
Capek, Petr
Bell, Sheryl L.
Hofmockel, Kirsten S.
Incomplete cell disruption of resistant microbes
title Incomplete cell disruption of resistant microbes
title_full Incomplete cell disruption of resistant microbes
title_fullStr Incomplete cell disruption of resistant microbes
title_full_unstemmed Incomplete cell disruption of resistant microbes
title_short Incomplete cell disruption of resistant microbes
title_sort incomplete cell disruption of resistant microbes
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6449382/
https://www.ncbi.nlm.nih.gov/pubmed/30948770
http://dx.doi.org/10.1038/s41598-019-42188-9
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