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Indirect DNA extraction method suitable for acidic soil with high clay content

DNA extraction is an essential procedure when investigating microbial communities in environmental samples by sequencing technologies. High clay soils can be problematic as DNA adsorbs to the clay particles and can thereby be preserved from lysed, non-viable cells for a substantial period of time. I...

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Autores principales: Högfors-Rönnholm, Eva, Christel, Stephan, Engblom, Sten, Dopson, Mark
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6314954/
https://www.ncbi.nlm.nih.gov/pubmed/30622912
http://dx.doi.org/10.1016/j.mex.2018.02.005
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author Högfors-Rönnholm, Eva
Christel, Stephan
Engblom, Sten
Dopson, Mark
author_facet Högfors-Rönnholm, Eva
Christel, Stephan
Engblom, Sten
Dopson, Mark
author_sort Högfors-Rönnholm, Eva
collection PubMed
description DNA extraction is an essential procedure when investigating microbial communities in environmental samples by sequencing technologies. High clay soils can be problematic as DNA adsorbs to the clay particles and can thereby be preserved from lysed, non-viable cells for a substantial period of time. In order to accurately estimate the intact and living microbial community in the soil, extracellular DNA from dead, remnant bacterial cells needs to be removed prior to DNA extraction. One possibility is to use a sodium phosphate buffer to release both extracellular DNA and bacterial cells from the clay particles. After removing the extracellular DNA by centrifugation, the remaining viable cells can be harvested and DNA extracted. The described method is a modification of a procedure for separating extracellular DNA and bacterial cells from acidic clay soils. • The modified method increases bacterial cell yields from acidic clay soils, such as acid sulfate soil. • The modified method eliminates some steps from the original method, as only DNA from intact bacterial cells is required. • The indirect DNA extraction method increases the workload compared to standard direct extraction methods, but subsequent downstream analyses will give a more representative picture of the viable microbial community composition in the soil.
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spelling pubmed-63149542019-01-08 Indirect DNA extraction method suitable for acidic soil with high clay content Högfors-Rönnholm, Eva Christel, Stephan Engblom, Sten Dopson, Mark MethodsX Agricultural and Biological Science DNA extraction is an essential procedure when investigating microbial communities in environmental samples by sequencing technologies. High clay soils can be problematic as DNA adsorbs to the clay particles and can thereby be preserved from lysed, non-viable cells for a substantial period of time. In order to accurately estimate the intact and living microbial community in the soil, extracellular DNA from dead, remnant bacterial cells needs to be removed prior to DNA extraction. One possibility is to use a sodium phosphate buffer to release both extracellular DNA and bacterial cells from the clay particles. After removing the extracellular DNA by centrifugation, the remaining viable cells can be harvested and DNA extracted. The described method is a modification of a procedure for separating extracellular DNA and bacterial cells from acidic clay soils. • The modified method increases bacterial cell yields from acidic clay soils, such as acid sulfate soil. • The modified method eliminates some steps from the original method, as only DNA from intact bacterial cells is required. • The indirect DNA extraction method increases the workload compared to standard direct extraction methods, but subsequent downstream analyses will give a more representative picture of the viable microbial community composition in the soil. Elsevier 2018-02-22 /pmc/articles/PMC6314954/ /pubmed/30622912 http://dx.doi.org/10.1016/j.mex.2018.02.005 Text en © 2018 The Authors http://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 Agricultural and Biological Science
Högfors-Rönnholm, Eva
Christel, Stephan
Engblom, Sten
Dopson, Mark
Indirect DNA extraction method suitable for acidic soil with high clay content
title Indirect DNA extraction method suitable for acidic soil with high clay content
title_full Indirect DNA extraction method suitable for acidic soil with high clay content
title_fullStr Indirect DNA extraction method suitable for acidic soil with high clay content
title_full_unstemmed Indirect DNA extraction method suitable for acidic soil with high clay content
title_short Indirect DNA extraction method suitable for acidic soil with high clay content
title_sort indirect dna extraction method suitable for acidic soil with high clay content
topic Agricultural and Biological Science
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6314954/
https://www.ncbi.nlm.nih.gov/pubmed/30622912
http://dx.doi.org/10.1016/j.mex.2018.02.005
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