PacBIO sequencing of Candidatus Pelagibacter giovannonii strain NP1: isolate genome
Cells were filtered onto sterile 0.2-mm Supor-200 polyethersulfone filters when cultures reached late exponential phase. DNA was then extracted and purity was verified by terminal restriction fragment length polymorphism analysis. Remaining DNA was purified on a single PowerClean DNA clean-up column (MO Bio Laboratories, Carlsbad, CA, USA). High-quality, purified DNA was recovered and submitted for genome sequencing. A PacBio Single-Molecule Real-Time (SMRT) 10kb library was constructed and sequenced using a single SMRT cell on the PacBio RSII platform to obtain 984 Mbp of DNA (raw reads). De-novo assembly of the genome from reads was conducted using Hierarchical Genome Assembly Process 4 (HGAP4). Through the HGAP4 pipeline, reads were preassembled, assembled using Celera, and polished with Quiver. The SMRT Analysis pipeline produced a single linear contig with overlapping ends and mean coverage of 697X. Genome ends were aligned and trimmed in Geneious v11.1.5 to produce a single 1.37Mbp chromosome.