title: Agrosoddy-podzolic soil, treated with benomyl (10x application rate), 14 days, bacterial/archaeal 16S
design: Soil samples were extracted with a micro-drill from microcosms, so as to collect the soil layer completely to the very bottom. Then the soil samples collected in this way were placed in 1.5 ml test tubes and frozen. Isolation of total DNA was carried out from 0.5 g samples of soil using the FastDNA SPIN Kit for Soil (MP Biomedicals, CA USA). Amplification of the variable region V4 of the 16S rRNA gene was carried out in one round using forward and reverse primers 515F (5-GTGCCAGCMGCCGCGGTAA-3) and 806R (5-GGACTACHVGGGTWTCTAAT-3) (Fadrosh et.al., 2014) with dual index sample multiplexing. These primers are specific to both bacteria and archaea. Amplification of the variable region ITS-1 was carried out using the following primers: BITS (5CTACCTGCGGARGGATCA3) and B58S3 (5 GAGATCCRTTGYTRAAAGTT3) (Mukherjee et.al., 2014). The PCR products were purified using the Cleanup Mini Kit (Evrogen, Russia) for DNA isolation from reaction mixtures. The concentration of the obtained libraries of 16S rRNA and ITS-1 in solution was measured on a Qubit fluorometer (Invitrogen, USA) using a Quant-iT dsDNA High-Sensitivity Assay Kit. The purified amplicons were mixed equimolarly in accordance with the concentrations obtained. The quality of the resulting library prepared for sequencing was assessed with agarose gel electrophoresis. Further sample preparation and sequencing of the pooled sample was carried out using a MiSeq sequencer (Illumina, USA) and the MiSeq Reagent Kit v2 (500 cycles). Forward and reverse sequences were pre-merged before the further bioinformatics analysis.