Cargando…

A simplified protocol for DNA extraction from FTA cards for faecal microbiome studies

As metagenomic studies continue to increase in size and complexity, they are often required to incorporate data from geographically isolated locations or longitudinal time samples. This represents a technical challenge, given that many of the commonly used methods used for sample collection, storage...

Descripción completa

Detalles Bibliográficos
Autores principales: Bolt Botnen, Amanda, Bjørnsen, Mads Bjørn, Alberdi, Antton, Gilbert, M.Thomas P., Aizpurua, Ostaizka
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9868478/
https://www.ncbi.nlm.nih.gov/pubmed/36699263
http://dx.doi.org/10.1016/j.heliyon.2023.e12861
_version_ 1784876547289645056
author Bolt Botnen, Amanda
Bjørnsen, Mads Bjørn
Alberdi, Antton
Gilbert, M.Thomas P.
Aizpurua, Ostaizka
author_facet Bolt Botnen, Amanda
Bjørnsen, Mads Bjørn
Alberdi, Antton
Gilbert, M.Thomas P.
Aizpurua, Ostaizka
author_sort Bolt Botnen, Amanda
collection PubMed
description As metagenomic studies continue to increase in size and complexity, they are often required to incorporate data from geographically isolated locations or longitudinal time samples. This represents a technical challenge, given that many of the commonly used methods used for sample collection, storage, and DNA extraction are sensitive to differences related to the time, storage and chemistry involved. FTA cards have been previously proposed as a simple, reliable and cost-efficient method for the preservation of animal faecal microbiomes. In this study, we report a simplified extraction methodology for recovering microbiome DNA from faeces stored on FTA cards and compare its performance to a common alternative means of characterising such microbiomes; namely, immediate freezing of the faeces followed by DNA extraction using the Qiagen PowerSoil DNA isolation kit. Our results show that overall the application of our simplified DNA extraction methodology yields microbial community results that have higher diversity and an expanded core microbiome than that found using the PowerSoil methodology. This suggests that the FTA card extraction method presented here is a viable alternative for metagenomic studies using faecal material when traditional freeze-based storage methods are not feasible.
format Online
Article
Text
id pubmed-9868478
institution National Center for Biotechnology Information
language English
publishDate 2023
publisher Elsevier
record_format MEDLINE/PubMed
spelling pubmed-98684782023-01-24 A simplified protocol for DNA extraction from FTA cards for faecal microbiome studies Bolt Botnen, Amanda Bjørnsen, Mads Bjørn Alberdi, Antton Gilbert, M.Thomas P. Aizpurua, Ostaizka Heliyon Research Article As metagenomic studies continue to increase in size and complexity, they are often required to incorporate data from geographically isolated locations or longitudinal time samples. This represents a technical challenge, given that many of the commonly used methods used for sample collection, storage, and DNA extraction are sensitive to differences related to the time, storage and chemistry involved. FTA cards have been previously proposed as a simple, reliable and cost-efficient method for the preservation of animal faecal microbiomes. In this study, we report a simplified extraction methodology for recovering microbiome DNA from faeces stored on FTA cards and compare its performance to a common alternative means of characterising such microbiomes; namely, immediate freezing of the faeces followed by DNA extraction using the Qiagen PowerSoil DNA isolation kit. Our results show that overall the application of our simplified DNA extraction methodology yields microbial community results that have higher diversity and an expanded core microbiome than that found using the PowerSoil methodology. This suggests that the FTA card extraction method presented here is a viable alternative for metagenomic studies using faecal material when traditional freeze-based storage methods are not feasible. Elsevier 2023-01-07 /pmc/articles/PMC9868478/ /pubmed/36699263 http://dx.doi.org/10.1016/j.heliyon.2023.e12861 Text en © 2023 The Authors https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Research Article
Bolt Botnen, Amanda
Bjørnsen, Mads Bjørn
Alberdi, Antton
Gilbert, M.Thomas P.
Aizpurua, Ostaizka
A simplified protocol for DNA extraction from FTA cards for faecal microbiome studies
title A simplified protocol for DNA extraction from FTA cards for faecal microbiome studies
title_full A simplified protocol for DNA extraction from FTA cards for faecal microbiome studies
title_fullStr A simplified protocol for DNA extraction from FTA cards for faecal microbiome studies
title_full_unstemmed A simplified protocol for DNA extraction from FTA cards for faecal microbiome studies
title_short A simplified protocol for DNA extraction from FTA cards for faecal microbiome studies
title_sort simplified protocol for dna extraction from fta cards for faecal microbiome studies
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9868478/
https://www.ncbi.nlm.nih.gov/pubmed/36699263
http://dx.doi.org/10.1016/j.heliyon.2023.e12861
work_keys_str_mv AT boltbotnenamanda asimplifiedprotocolfordnaextractionfromftacardsforfaecalmicrobiomestudies
AT bjørnsenmadsbjørn asimplifiedprotocolfordnaextractionfromftacardsforfaecalmicrobiomestudies
AT alberdiantton asimplifiedprotocolfordnaextractionfromftacardsforfaecalmicrobiomestudies
AT gilbertmthomasp asimplifiedprotocolfordnaextractionfromftacardsforfaecalmicrobiomestudies
AT aizpuruaostaizka asimplifiedprotocolfordnaextractionfromftacardsforfaecalmicrobiomestudies
AT boltbotnenamanda simplifiedprotocolfordnaextractionfromftacardsforfaecalmicrobiomestudies
AT bjørnsenmadsbjørn simplifiedprotocolfordnaextractionfromftacardsforfaecalmicrobiomestudies
AT alberdiantton simplifiedprotocolfordnaextractionfromftacardsforfaecalmicrobiomestudies
AT gilbertmthomasp simplifiedprotocolfordnaextractionfromftacardsforfaecalmicrobiomestudies
AT aizpuruaostaizka simplifiedprotocolfordnaextractionfromftacardsforfaecalmicrobiomestudies