V4 16S rRNA amplicon sequencing of marine eDNA from the Levantine basin
DNA was extracted from water samples collected on 0.2 micron Supor filters with the PowerWater kit (Qiagen, USA), using the FastPrep-24 Classic (MP Biomedicals, USA) bead-beating to disrupt the cells (2 cycles at 5.5 m sec-1, with a 5 min interval).  The V4 region (~ 300 bp) of the 16S rRNA gene was amplified from the DNA (~50 ng) using the 515Fc/806Rc primers amended with relevant tags (Apprill et al., 2015; Parada et al., 2016). Following first PCR, library preparation and sequencing were performed with relevant protocols.