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
Peer reviewed, Journal article
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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.