PacBio Sequencing of Salinibacter ruber Strain M31T
DNA samples (about 2mg) were sheared to an average size of ~10 kb using the G-tube protocol (Covaris, MA). DNA libraries were prepared using a SMRTbell express template prep kit 2.0 (100-938-900, Pacific Bioscience, CA) and ligated with barcoded hairpin lbc--lbc adapters for multiplex sequencing on a Sequel 2 (SQ2) instrument. Incompletely formed SMRTbell templates were removed by digestion with a combination of exonuclease III and exonuclease VII (NEB, MA).  The qualification and quantification of the SMRTbell libraries were made on a Qubit fluorimeter (Invitrogen, OR) and a 2100 Bioanalyzer (Agilent Technologies, CA).  Size-selected separation of SMRTbell libraries on gel-based Blue Pippin instrument (Sage Science, Beverly, MA) was performed to remove impurities and improve genome assembly results.