16S rRNA amplicon sequencing of negative control
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 V4: [515FB] GTGYCAGCMGCCGCGGTAAand [806RB] GGACTACNVGGGTWTCTAAT (Apprill et al., 2015) with the following conditions were pooled: Initial denaturation at 95 C for 2 min, 30 cycles of amplification (95 C for 15 s, 55 C for 15 s, 72 C for 50s) and a final elongation at 72 C for 5 min. After the second PCR, sequencing was performed using a MiSeq Reagent kit v3 (Illumina, USA)
MQ201102-227