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Inter-laboratory testing of the effect of DNA blocking reagent G2 on DNA extraction from low-biomass clay samples

Here we show that a commercial blocking reagent (G2) based on modified eukaryotic DNA significantly improved DNA extraction efficiency. We subjected G2 to an inter-laboratory testing, where DNA was extracted from the same clay subsoil using the same batch of kits. The inter-laboratory extraction cam...

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Autores principales: Jacobsen, Carsten Suhr, Nielsen, Tue Kjærgaard, Vester, Jan Kjølhede, Stougaard, Peter, Nielsen, Jeppe Lund, Voriskova, Jana, Winding, Anne, Baldrian, Petr, Liu, Binbin, Frostegård, Åsa, Pedersen, Dorthe, Tveit, Alexander Tøsdal, Svenning, Mette Marianne, Tebbe, Christoph C., Øvreås, Lise, Jakobsen, Pia Bach, Blazewicz, Steven J., Hubablek, Valerie, Bertilsson, Stefan, Hansen, Lars Hestbjerg, Cary, S. Craig, Holben, William E., Ekelund, Flemming, Bælum, Jacob
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5890260/
https://www.ncbi.nlm.nih.gov/pubmed/29632323
http://dx.doi.org/10.1038/s41598-018-24082-y
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author Jacobsen, Carsten Suhr
Nielsen, Tue Kjærgaard
Vester, Jan Kjølhede
Stougaard, Peter
Nielsen, Jeppe Lund
Voriskova, Jana
Winding, Anne
Baldrian, Petr
Liu, Binbin
Frostegård, Åsa
Pedersen, Dorthe
Tveit, Alexander Tøsdal
Svenning, Mette Marianne
Tebbe, Christoph C.
Øvreås, Lise
Jakobsen, Pia Bach
Blazewicz, Steven J.
Hubablek, Valerie
Bertilsson, Stefan
Hansen, Lars Hestbjerg
Cary, S. Craig
Holben, William E.
Ekelund, Flemming
Bælum, Jacob
author_facet Jacobsen, Carsten Suhr
Nielsen, Tue Kjærgaard
Vester, Jan Kjølhede
Stougaard, Peter
Nielsen, Jeppe Lund
Voriskova, Jana
Winding, Anne
Baldrian, Petr
Liu, Binbin
Frostegård, Åsa
Pedersen, Dorthe
Tveit, Alexander Tøsdal
Svenning, Mette Marianne
Tebbe, Christoph C.
Øvreås, Lise
Jakobsen, Pia Bach
Blazewicz, Steven J.
Hubablek, Valerie
Bertilsson, Stefan
Hansen, Lars Hestbjerg
Cary, S. Craig
Holben, William E.
Ekelund, Flemming
Bælum, Jacob
author_sort Jacobsen, Carsten Suhr
collection PubMed
description Here we show that a commercial blocking reagent (G2) based on modified eukaryotic DNA significantly improved DNA extraction efficiency. We subjected G2 to an inter-laboratory testing, where DNA was extracted from the same clay subsoil using the same batch of kits. The inter-laboratory extraction campaign revealed large variation among the participating laboratories, but the reagent increased the number of PCR-amplified16S rRNA genes recovered from biomass naturally present in the soils by one log unit. An extensive sequencing approach demonstrated that the blocking reagent was free of contaminating DNA, and may therefore also be used in metagenomics studies that require direct sequencing.
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spelling pubmed-58902602018-04-13 Inter-laboratory testing of the effect of DNA blocking reagent G2 on DNA extraction from low-biomass clay samples Jacobsen, Carsten Suhr Nielsen, Tue Kjærgaard Vester, Jan Kjølhede Stougaard, Peter Nielsen, Jeppe Lund Voriskova, Jana Winding, Anne Baldrian, Petr Liu, Binbin Frostegård, Åsa Pedersen, Dorthe Tveit, Alexander Tøsdal Svenning, Mette Marianne Tebbe, Christoph C. Øvreås, Lise Jakobsen, Pia Bach Blazewicz, Steven J. Hubablek, Valerie Bertilsson, Stefan Hansen, Lars Hestbjerg Cary, S. Craig Holben, William E. Ekelund, Flemming Bælum, Jacob Sci Rep Article Here we show that a commercial blocking reagent (G2) based on modified eukaryotic DNA significantly improved DNA extraction efficiency. We subjected G2 to an inter-laboratory testing, where DNA was extracted from the same clay subsoil using the same batch of kits. The inter-laboratory extraction campaign revealed large variation among the participating laboratories, but the reagent increased the number of PCR-amplified16S rRNA genes recovered from biomass naturally present in the soils by one log unit. An extensive sequencing approach demonstrated that the blocking reagent was free of contaminating DNA, and may therefore also be used in metagenomics studies that require direct sequencing. Nature Publishing Group UK 2018-04-09 /pmc/articles/PMC5890260/ /pubmed/29632323 http://dx.doi.org/10.1038/s41598-018-24082-y Text en © The Author(s) 2018 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
Jacobsen, Carsten Suhr
Nielsen, Tue Kjærgaard
Vester, Jan Kjølhede
Stougaard, Peter
Nielsen, Jeppe Lund
Voriskova, Jana
Winding, Anne
Baldrian, Petr
Liu, Binbin
Frostegård, Åsa
Pedersen, Dorthe
Tveit, Alexander Tøsdal
Svenning, Mette Marianne
Tebbe, Christoph C.
Øvreås, Lise
Jakobsen, Pia Bach
Blazewicz, Steven J.
Hubablek, Valerie
Bertilsson, Stefan
Hansen, Lars Hestbjerg
Cary, S. Craig
Holben, William E.
Ekelund, Flemming
Bælum, Jacob
Inter-laboratory testing of the effect of DNA blocking reagent G2 on DNA extraction from low-biomass clay samples
title Inter-laboratory testing of the effect of DNA blocking reagent G2 on DNA extraction from low-biomass clay samples
title_full Inter-laboratory testing of the effect of DNA blocking reagent G2 on DNA extraction from low-biomass clay samples
title_fullStr Inter-laboratory testing of the effect of DNA blocking reagent G2 on DNA extraction from low-biomass clay samples
title_full_unstemmed Inter-laboratory testing of the effect of DNA blocking reagent G2 on DNA extraction from low-biomass clay samples
title_short Inter-laboratory testing of the effect of DNA blocking reagent G2 on DNA extraction from low-biomass clay samples
title_sort inter-laboratory testing of the effect of dna blocking reagent g2 on dna extraction from low-biomass clay samples
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5890260/
https://www.ncbi.nlm.nih.gov/pubmed/29632323
http://dx.doi.org/10.1038/s41598-018-24082-y
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