title: Enrichment and isolation of aerobic and anaerobic hydrocarbon-degrading bacteria from the multicontaminated marine sediment of the Mar Piccolo of Taranto (Italy)
design: 4 ng of DNA was used for the 16S rDNA gene sequencing targeting the V1V3 variable region. PCR reactions for the library construction were performed in 25 L total volume containing Phusion Master Mix High Fidelity (Thermo Fisher Scientific, United States) and 0.5 M final concentration of the library adaptors with 27F and 534R primers. All PCR reactions were run in duplicate and pooled afterward. The amplicon libraries were purified using the Agencourt AMpureXP-beads (Beckmann Coulter, United States) and the concentrations were measured with Qubit 3.0 fluorometer (Thermo Fisher Scientific, United States). Purified libraries were pooled in equimolar concentrations and diluted to 4 nM. Phix control was added at 10% in the pooled libraries. Samples were paired-end sequenced (2  301 bp) on a MiSeq (Illumina, United States) instrument using a MiSeq Reagent kit v3, 600 cycles (Illumina, United States) following the standard guidelines