Dataset: 16S rRNA gene clones from NE Pacific bacterioplankton
Deployment: TN268

Nearly full-length 16S rRNA gene sequences were obtained for frequently occuring lineages by selecting and purifying a subset of clones using the Qiagen plasmid mini-prep kit (QIAGEN, Germantown, MD). Sequencing was performed using the following primer pairs (M13R-, 519F (5′-CAGC(A/G)GCCGCGGTAATAC-3′), 338F (5′-ACTCCTACGGGAGGCAGC-3′), and 926R (5′-CCGTCAATTC(A/C)TTT (A/G)AGTTT-3′)) by GeneWiz, Inc. (LaJolla, CA). Sequences were assembled with the CAP3 Sequence Assembly Program (Huang and Madan 1999) and annotated by using the Bayesian method of (Wang et al 2007), which contained a custom training set augmented with marine environmental clades (Iverson et al 2012), and by using the SILVA least common ancestor classification tool (Pruesse et al 2007). In silico restriction analysis and other DNA sequence manipulation operations were performed with the Sequence Manipulation Suite (Stothard 2000). Restriction fragments were verified for a subset of clones to confirm predicted fragment sizes. Bacterial 16S rRNA sequences generated during this study were submitted to Genbank (accession numbers KC522839- KC522949 -- See Data).