title: metagenome of paddy wetland exposed to heavy metal and pesticides
design: The samples were then diluted to 1ng/L in Low TE and subjected to library preparation workflow for the 16S rRNA gene V3-V4 region using the Illumina demonstrated protocol for 16S Metagenomic sequencing using a phased primer strategy. The protocol uses a phased primer strategy as outlined by Fadrosh et al. (2014) . Indexed libraries were purified and pooled at equimolar concentrations to a concentration of 10nM. The pool was further validated using an Agilent Tapestation 4200 high sensitivity D1000 screentape (Agilent) and quantified using the qubit fluorometer high sensitivity ds DNA kit (Thermo Fisher Scientific). The pooled library was sequenced on an Illumina MiSeq instrument using the 2x249 base chemistry of the v2 reagent kit as per manufacturers instructions. The reads produced were demultiplexed on board the sequencer and raw fastq files were generated.