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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; Frostegard, Åsa; Pedersen, Dorthe; Tveit, Alexander Tøsdal; Svenning, Mette Marianne; Tebbe, Christoph C.; Ovreas, Lise Øvreås; Jakobsen, Pia Bach; Blazewicz, Steven J.; Hubablek, Valerie; Bertilsson, Stefan; Hansen, Lars Hestbjerg; Cary, S. Craig; Holben, William E.; Ekelund, Flemming; Baelum, Jacob
Abstract
Here we show that a commercial blocking reagent (G2) based on modifed eukaryotic DNA signifcantly improved DNA extraction efciency. 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-amplifed16S 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.
Type
Journal Article
Type of thesis
Series
Citation
Jacobsen, C. S., Nielsen, T. K., Vester, J. K., Stougaard, P., Nielsen, J. L., Voriskova, J., … Baelum, J. (2018). Inter-laboratory testing of the effect of DNA blocking reagent G2 on DNA extraction from low-biomass clay samples. Scientific Reports, 8. https://doi.org/10.1038/s41598-018-24082-y
Date
2018-04-09
Publisher
Nature Publishing Group
Degree
Supervisors
Rights
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/.