PacBio sequencing of Wolbachia wAlbB from Aedes aegypti cell line Aa23
High molecular weight (larger than 60 kb in size), good quality (DIN > 8.2) genomic DNA was extracted from the cell-free Wolbachia wAlbB obtained from spent culture media. Genomic DNA was fragmented using a Megaruptor 2 device (Diagenode) targeting a sheared size of 20 kb.  Sheared DNA was purified with AMPure PB beads and 2ug were used to construct a SMRTbell library according to PacBio library construction guidelines. Briefly, sheared DNA was repaired using the NEB Next FFPE DNA enzyme mix, followed by end-repair to generate blunt ends.  Following purification using AMPure PB magnetic beads, PacBio universal hairpin adaptors were ligated to the DNA to produce SMRTbell libraries. After adaptor removal and library clean up, the library was loaded on a PacBio RSII system using a MagBead binding kit and sequenced with a 300 minutes collection time.