Dataset: TetV-1 genome annotation

Note: The detailed protocols are described in Schvarcz and Steward, 2018. 

Shoreline sampling was carried out on September 2nd, 2010 in the coastal surface water of Kāneʻohe Bay, Oʻahu, Hawaiʻi.  Seawater (175 L) was filtered (0.8 µm polycarbonate track-etched), and particles in the filtrate concentrated by tangential flow filtration (TFF; 30 kDA molecular weight cut-off, regenerated cellulose). This was then used to challenge a culture of Tetraselmis species isolated previously from the same water. Lysate was serially diluted and used to challenge a new culture for several rounds using a dilution-to-extinction approach to ensure clonality of the virus. A large batch of lysate (40 L of culture) was filtered (0.45 PVDF over a Whatman glass-fiber GF/C). Viruses in the filtrate were concentrated by TFF (30 kDa) to 350 mL volume, further concentrated by polyethylene glycol precipitation, and purified in a CsCl buoyant density gradient. DNA was extracted from the virus peak in the gradient by buffer exchange into TE, treatment with hot SDS and proteinase K, followed by sequential selective precipitation of proteins (salting out), then DNA (alcohol precipitation) using the MasterPure DNA Purification Kit (Epicentre).

DNA was sequenced using PacBio P6-C4 chemistry platform.  The complete genome sequence was deposited in GenBank (Clark, 2015) with accession number KY322437 (https://www.ncbi.nlm.nih.gov/nuccore/KY322437).  The gene annotations were published as Supplementary Dataset S1 in Schvarcz & Steward, 2018.