Dataset: 16S microbiome metadata collected from shallow artificial reef sponges and seawater in the Florida Keys, USA from Apr 2021 to Aug 2021

Location
Samples were collected on shallow reefs (<10m) in the Florida Keys, USA. The temporary artificial reef was constructed adjacent to patch reefs within the Looe Key special preservation area. At the conclusion of the experiment, the reef was completely dismantled. The environment was well mixed by wave action within minimal tidal influence.

Collection and Analysis

Seawater samples were collected via VacuSIP and filtered through 0.2 µm membranes with a peristaltic pump to collect samples of ambient microbes. Total genomic DNA was extracted from cross sections of sponge tissue (~0.25 grams) and ½ of seawater filter membranes using Qiagen PowerSoil Powerlyzer extraction kit and following the manufacturers protocol. After DNA extraction, polymerase chain reaction (PCR) was performed following the 16S Illumina Amplicon protocol of the Earth Microbiome project (Caporaso, 2018) with barcoded 16S rRNA primers (515F and 806R; (Apprill et al. 2015, Parada et al. 2016) and PCR products were cleaned using AMPure beads (Beckman Coulter). DNA concentration in cleaned products was measured using a Qubit fluorometer (Qubit). The concentration of each sample was diluted to 4 nM and then all samples were pooled in equal volumes. Pooled samples were then sequenced on an Illumina MiSeq platform following standard Illumina protocols for sample preparation and loading except that custom sequencing primers from the EMP protocol were used. A 500 cycle V2 chemistry MiSeq kit was used during sequencing, which yielded paired-end 250 base pair (bp) amplicons.