Bacterial 16S rRNA genes from interface layer of unspiked control column, replicate a
A two-layer column study with/without phenanthrene spike present in the upper soil layer was conducted under water-unsaturated flow conditions in order to study the effect of mobile organic matter on soil bacteria. After irrigation, total community DNA (TC-DNA) was extracted from three different soil depths. Purified TC-DNA was used as target to amplify the V3-V4 region of 16S rRNA genes with primers 338F/802R fused to Roche-454 A and B Titanium sequencing adapters and an eight-base barcode sequence in adaptor A. Library preparation and sequencing were done on a 454 Genome Sequencer FLX Titanium platform according to standard 454 protocols (Roche 454 Life Sciences, Branford, CT) by Biocant, Genoinseq unit (Cantanhede, Portugal).