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

Carsten Suhr Jacobsen*, Tue Kjaergaard Nielsen, Jan Kjolhede Vester, Peter Stougaard, Jeppe Lund Nielsen, Jana Voriskova, Anne Winding, Petr Baldrian, Binbin Liu, Asa Frostegard, Dorthe Pedersen, Alexander Tosdal Tveit, Mette Marianne Svenning, Christoph C. Tebbe, Lise Ovreas, Pia Bach Jakobsen, Steven J. Blazewicz, Valerie Hubablek, Stefan Bertilsson, Lars Hestbjerg HansenS. Craig Cary, William E. Holben, Flemming Ekelund, Jacob Baelum

*Corresponding author af dette arbejde

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Abstract

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.

OriginalsprogEngelsk
Artikelnummer5711
TidsskriftScientific Reports
Vol/bind8
Nummer1
Antal sider6
ISSN2045-2322
DOI
StatusUdgivet - 9 apr. 2018

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