title: Treatment of wastewater from the soluble coffee industry in an anaerobic sequencing batch reactor with immobilized biomass (AnSBBR) followed by physico-chemical processes of coagulation, flocculation, and sedimentation
design: DNA was extracted using the FastDNA SPIN for Soil DNA Extraction kit (MP, Bi-omedicals) according to the manufacturer's recommendations. DNA quantification (ng/?L) and purity (ratio 260/280 = 1.8 - 2.0) were performed using a Nanodrop 2000 spec-trophotometer (ThermoFisher Scientific). The integrity of the DNA fragment was analyzed by 0.8% agarose gel electrophoresis. DNA samples were sent to the company NGS Solu-es Genmicas, located in the city of Piracicaba - SP (www.ngsgenomica.com.br), for sequencing of the 16S rRNA gene. Taxonomic characterization was performed based on DNA amplification, using specific primers for the V3-V4 (341F/785R) and V4 (Ar915aF-Ar1386R) regions of the 16S rRNA gene (KITTELMANN et al., 2013; KLINDWORTH et al., 2013). Sequencing was performed on the Illumina MiSeq platform, configured for paired-end reads (2x300 bp), ensuring an average coverage of 50,000 paired-end reads per sample.