16S rRNA V3-V4 amplicon sequencing of freshwater metagenome from Rio Abobral, Brazilian Pantanal
Water samples (3 L) were filtered through 0.22 um sterile membranes. DNA was extracted using the DNeasy PowerWater Kit (QIAGEN). The V3-V4 region of the 16S rRNA gene was amplified using primers 341F (CCTACGGGNGGCWGCAG) and 785R (GACTACHVGGGTATCTAATCC). Libraries were prepared with Native Barcoding Kit 24 V14 (SQK-NBD114.24, Oxford Nanopore Technologies) and sequenced on a MinION Mk1B device with FLO-MIN114 (R10.4.1) flow cells. Basecalling and demultiplexing were performed using MinKNOW.