Cargando…
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...
Autores principales: | , , , , , , , , , , , , , , , , , , , , , , , |
---|---|
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 |
_version_ | 1783312839251853312 |
---|---|
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. |
format | Online Article Text |
id | pubmed-5890260 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT jacobsencarstensuhr interlaboratorytestingoftheeffectofdnablockingreagentg2ondnaextractionfromlowbiomassclaysamples AT nielsentuekjærgaard interlaboratorytestingoftheeffectofdnablockingreagentg2ondnaextractionfromlowbiomassclaysamples AT vesterjankjølhede interlaboratorytestingoftheeffectofdnablockingreagentg2ondnaextractionfromlowbiomassclaysamples AT stougaardpeter interlaboratorytestingoftheeffectofdnablockingreagentg2ondnaextractionfromlowbiomassclaysamples AT nielsenjeppelund interlaboratorytestingoftheeffectofdnablockingreagentg2ondnaextractionfromlowbiomassclaysamples AT voriskovajana interlaboratorytestingoftheeffectofdnablockingreagentg2ondnaextractionfromlowbiomassclaysamples AT windinganne interlaboratorytestingoftheeffectofdnablockingreagentg2ondnaextractionfromlowbiomassclaysamples AT baldrianpetr interlaboratorytestingoftheeffectofdnablockingreagentg2ondnaextractionfromlowbiomassclaysamples AT liubinbin interlaboratorytestingoftheeffectofdnablockingreagentg2ondnaextractionfromlowbiomassclaysamples AT frostegardasa interlaboratorytestingoftheeffectofdnablockingreagentg2ondnaextractionfromlowbiomassclaysamples AT pedersendorthe interlaboratorytestingoftheeffectofdnablockingreagentg2ondnaextractionfromlowbiomassclaysamples AT tveitalexandertøsdal interlaboratorytestingoftheeffectofdnablockingreagentg2ondnaextractionfromlowbiomassclaysamples AT svenningmettemarianne interlaboratorytestingoftheeffectofdnablockingreagentg2ondnaextractionfromlowbiomassclaysamples AT tebbechristophc interlaboratorytestingoftheeffectofdnablockingreagentg2ondnaextractionfromlowbiomassclaysamples AT øvreaslise interlaboratorytestingoftheeffectofdnablockingreagentg2ondnaextractionfromlowbiomassclaysamples AT jakobsenpiabach interlaboratorytestingoftheeffectofdnablockingreagentg2ondnaextractionfromlowbiomassclaysamples AT blazewiczstevenj interlaboratorytestingoftheeffectofdnablockingreagentg2ondnaextractionfromlowbiomassclaysamples AT hubablekvalerie interlaboratorytestingoftheeffectofdnablockingreagentg2ondnaextractionfromlowbiomassclaysamples AT bertilssonstefan interlaboratorytestingoftheeffectofdnablockingreagentg2ondnaextractionfromlowbiomassclaysamples AT hansenlarshestbjerg interlaboratorytestingoftheeffectofdnablockingreagentg2ondnaextractionfromlowbiomassclaysamples AT caryscraig interlaboratorytestingoftheeffectofdnablockingreagentg2ondnaextractionfromlowbiomassclaysamples AT holbenwilliame interlaboratorytestingoftheeffectofdnablockingreagentg2ondnaextractionfromlowbiomassclaysamples AT ekelundflemming interlaboratorytestingoftheeffectofdnablockingreagentg2ondnaextractionfromlowbiomassclaysamples AT bælumjacob interlaboratorytestingoftheeffectofdnablockingreagentg2ondnaextractionfromlowbiomassclaysamples |