title: 454 GS FLX Titanium sequencing; 454 sequencing of GS734 0p1µm filter fraction
design: 454 sequencing for use in comparative metagenomics of global surface oceans,                 GOS site GS734, 0p1µm filter fraction
library_construction_protocol: Environmental DNA (eDNA) was isolated from the impact filters and                 library construction was performed according to Rusch, et al. 2007. Briefly, shipboard frozen                 filters contained in the following storage buffer: 1X Tris-EDTA (TE) buffer (Invitrogen), 50mM                 EGTA and 50mM EDTA, were thawed to room temperature and aseptically cut for DNA extraction.                  Filter fragments were placed in 50ml conical tubes and lysis was performed as previously described.                  DNA was ultimately purified using a phenol/chlorofom extraction method followed by ethanol                  precipitation.                   eDNA was sheared and 454 adaptors ligated. Subsequently, AMPure bead purification was followed                  by emulsion PCR (emPCR) and sequenced on the Roche 454 Sequencing platform.