16S rRNA amplicon sequencing of a freshwater microbial community
Filtering of water samples and extraction of DNA using the standard protocol for Dneasy PowerWater Kit (Qiagen, Germany). Duplicate amplification of the 16S rRNA gene region 18 (bV18A): [8F] AGRGTTYGATYMTGGCTCAGand [1391R] GACGGGCGGTGWGTRCA. Initial denaturation at 98 C for 3 min, 25 cycles of amplification (98 C for 30 s, 62 C for 20 s,72 C for 2 min) and a final elongation at 72 C for 5 min. Sequencing was performed using a MinION R10.4.1 flowcell (Oxford Nanopore Technologies, UK)