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An efficient method for high molecular weight bacterial DNA extraction suitable for shotgun metagenomics from skin swabs

The human skin microbiome represents a variety of complex microbial ecosystems that play a key role in host health. Molecular methods to study these communities have been developed but have been largely limited to low-throughput quantification and short amplicon-based sequencing, providing limited f...

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Autores principales: Serghiou, Iliana R., Baker, Dave, Evans, Rhiannon, Dalby, Matthew J., Kiu, Raymond, Trampari, Eleftheria, Phillips, Sarah, Watt, Rachel, Atkinson, Thomas, Murphy, Barry, Hall, Lindsay J., Webber, Mark A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Microbiology Society 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10438817/
https://www.ncbi.nlm.nih.gov/pubmed/37428148
http://dx.doi.org/10.1099/mgen.0.001058
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author Serghiou, Iliana R.
Baker, Dave
Evans, Rhiannon
Dalby, Matthew J.
Kiu, Raymond
Trampari, Eleftheria
Phillips, Sarah
Watt, Rachel
Atkinson, Thomas
Murphy, Barry
Hall, Lindsay J.
Webber, Mark A.
author_facet Serghiou, Iliana R.
Baker, Dave
Evans, Rhiannon
Dalby, Matthew J.
Kiu, Raymond
Trampari, Eleftheria
Phillips, Sarah
Watt, Rachel
Atkinson, Thomas
Murphy, Barry
Hall, Lindsay J.
Webber, Mark A.
author_sort Serghiou, Iliana R.
collection PubMed
description The human skin microbiome represents a variety of complex microbial ecosystems that play a key role in host health. Molecular methods to study these communities have been developed but have been largely limited to low-throughput quantification and short amplicon-based sequencing, providing limited functional information about the communities present. Shotgun metagenomic sequencing has emerged as a preferred method for microbiome studies as it provides more comprehensive information about the species/strains present in a niche and the genes they encode. However, the relatively low bacterial biomass of skin, in comparison to other areas such as the gut microbiome, makes obtaining sufficient DNA for shotgun metagenomic sequencing challenging. Here we describe an optimised high-throughput method for extraction of high molecular weight DNA suitable for shotgun metagenomic sequencing. We validated the performance of the extraction method, and analysis pipeline on skin swabs collected from both adults and babies. The pipeline effectively characterised the bacterial skin microbiota with a cost and throughput suitable for larger longitudinal sets of samples. Application of this method will allow greater insights into community compositions and functional capabilities of the skin microbiome.
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spelling pubmed-104388172023-08-19 An efficient method for high molecular weight bacterial DNA extraction suitable for shotgun metagenomics from skin swabs Serghiou, Iliana R. Baker, Dave Evans, Rhiannon Dalby, Matthew J. Kiu, Raymond Trampari, Eleftheria Phillips, Sarah Watt, Rachel Atkinson, Thomas Murphy, Barry Hall, Lindsay J. Webber, Mark A. Microb Genom Methods The human skin microbiome represents a variety of complex microbial ecosystems that play a key role in host health. Molecular methods to study these communities have been developed but have been largely limited to low-throughput quantification and short amplicon-based sequencing, providing limited functional information about the communities present. Shotgun metagenomic sequencing has emerged as a preferred method for microbiome studies as it provides more comprehensive information about the species/strains present in a niche and the genes they encode. However, the relatively low bacterial biomass of skin, in comparison to other areas such as the gut microbiome, makes obtaining sufficient DNA for shotgun metagenomic sequencing challenging. Here we describe an optimised high-throughput method for extraction of high molecular weight DNA suitable for shotgun metagenomic sequencing. We validated the performance of the extraction method, and analysis pipeline on skin swabs collected from both adults and babies. The pipeline effectively characterised the bacterial skin microbiota with a cost and throughput suitable for larger longitudinal sets of samples. Application of this method will allow greater insights into community compositions and functional capabilities of the skin microbiome. Microbiology Society 2023-07-10 /pmc/articles/PMC10438817/ /pubmed/37428148 http://dx.doi.org/10.1099/mgen.0.001058 Text en © 2023 The Authors https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License. This article was made open access via a Publish and Read agreement between the Microbiology Society and the corresponding author’s institution.
spellingShingle Methods
Serghiou, Iliana R.
Baker, Dave
Evans, Rhiannon
Dalby, Matthew J.
Kiu, Raymond
Trampari, Eleftheria
Phillips, Sarah
Watt, Rachel
Atkinson, Thomas
Murphy, Barry
Hall, Lindsay J.
Webber, Mark A.
An efficient method for high molecular weight bacterial DNA extraction suitable for shotgun metagenomics from skin swabs
title An efficient method for high molecular weight bacterial DNA extraction suitable for shotgun metagenomics from skin swabs
title_full An efficient method for high molecular weight bacterial DNA extraction suitable for shotgun metagenomics from skin swabs
title_fullStr An efficient method for high molecular weight bacterial DNA extraction suitable for shotgun metagenomics from skin swabs
title_full_unstemmed An efficient method for high molecular weight bacterial DNA extraction suitable for shotgun metagenomics from skin swabs
title_short An efficient method for high molecular weight bacterial DNA extraction suitable for shotgun metagenomics from skin swabs
title_sort efficient method for high molecular weight bacterial dna extraction suitable for shotgun metagenomics from skin swabs
topic Methods
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10438817/
https://www.ncbi.nlm.nih.gov/pubmed/37428148
http://dx.doi.org/10.1099/mgen.0.001058
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