Amplicon sequencing of coral reef seawater over a diel and daily time-series
Seawater was filtered onto 0.22 um SUPOR DNA filters using a 60 mL syringe and DNA was extracted from the filters using a variation of the sucrose-lysis DNA extraction method. The universal v4 primers 515F and 806R (Apprill et. al, 2014; Parada et al., 2015) were used to amplify and barcode the v4 hypervariable region on the 16S rRNA gene of bacteria and archaea. Barcoded PCR products were pooled into a single library with equimolar ratios and sequenced using Illumina MiSeq sequencing.