Amplicon sequencing of environmental sample: water from Great Bay Estuary, Oyster River, NH
A 10ng/mL multiplexed sample was prepared for the Roche Genome Sequencer FLX System using Titanium Chemistry (454 Life Sciences, Branford, CT, USA). The DNA concentration for each sample was quantified using a NanoDrop 2000c (Thermo Scientific, Wilmington, DE, USA) and pooled with equal proportions of the twenty oyster and two water samples.  The pooled mixture was purified using the AMPure XP Purification Kit (Beckman Coulter Genomics, Danvers, MA, USA) by manufacturers protocols, with the final samples suspended in 20uL elution buffer EB from the MinElute PCR Purification Kit (Qiagen, Valencia, CA, USA).  The pooled tagged single-stranded pyrosequencing library underwent fusion PCR and pyrosequencing using a Roche 454 FLX Pyrosequencer (454 Life Sciences, Branford, CT, USA) according to the manufacturer instructions at the University of Illinois W.M. Keck Center High-Throughout DNA Sequencing Center.