Elba Pomonte reference site 2 - Bacteria
Total nucleic acids were extracted from 1 ml sediment using a Fast DNA Spin Kit For Soil (MP Biomedicals) and were purified using Wizard DNA Clean-Up (Promega). The quality was checked and the concentration was measured by spectrophotometry (Qubit 2.0 Fluorometer or Infinite 200 Nano Quant) and the aliquots stored at -20°C until further processing. PCR amplification of the bacterial 16S rRNA gene variable regions V3-V5 was done with standardized amounts of DNA using the primers 341F (5’-CCTACGGGAGGCAGCAG-3’) and 907R (5’-CCGTCAATTCMTTTGAGTTT-3’). Amplicon purification, library preparation and massively parallel tag sequencing using a 454 GS FLX+ sequencer (Roche) was carried out at the Research and Testing Laboratory (Lubbock, Texas).