16S rRNA gene amplicon sequencing of soil, groundwater, river sediment and river water in the region of Tuebingen, Germany.
More...16S rRNA gene amplicon sequencing of soil, groundwater, river sediment and river water in the region of Tuebingen, Germany. The samples were used to benchmark 16S rRNA gene amplicon sequencing analysis tools, namely Mothur, QIIME, QIIME2 and MEGAN.
Water samples were filtered (0.2 um). DNA extraction was performed following Lueders et al. (2004). Bacterial and archaeal 16S rRNA genes were amplified using universal primers 515f and 806r fused to Illumina adapters. Subsequent library preparation steps (Nextera, Illumina) and 250 bp paired-end sequencing with MiSeq (Illumina, San Diego, CA, USA) using v2 chemistry were performed by Microsynth AG (Switzerland). Less...