title: Deep Sea Sponge and Seawater_SW1
design: PCR amplicon libraries of the V5-V6 region of 16S rRNA genes were prepared from metagenomic DNA. Universal primers U789f (5’-TAGATACCCSSGTAGTCC-3’) and U1068r (5’-CTGACGRCRGCCATGC-3’), targeting both bacteria and archaea, were adapted for pyrosequencing by the addition of sequencing adapters and multiplex identifier (MID) sequences.  To minimise PCR bias three individual reactions were performed per template and equimolar amounts of PCR products from each of the three reactions were pooled for pyrosequencing. PCR products were purified using Qiagen PCR Purification Kit (Qiagen Ltd., UK) as per the manufacturer’s instructions. Barcoded samples were pooled and sequenced on a GS FLX Titanium platform.