16S amplicon metabarcoding of hydrothermal vent plume water: Cast 1, Bottle 12, Filter 2
Metagenomic DNA was extracted with the Macherey-Nagel NucleoMag DNA/RNA Water kit. DNA quantity was measured with the Qubit HS DNA assay. The V4 hypervariable region of the 16S SSU rRNA gene was targeted with the following prokaryote-specific forward and reverse primers, respectively: 5'-GTGYCAGCMGCCGCGGTAA-3' (Parada et al. 2016) and 5'-GGACTACNVGGGTWTCTAAT-3' (Apprill et al. 2015). Primers included linker sequence on their 5 ends to additional of Illumina-compatible dual-index adapter sequences. Sequencing was performed on a MiSeq using V3 reagents (2x300 bp).