Bacillus cereus 03BB102
Purified gDNA was sheared to a target size of 12-15 kbp in Covaris g-Tubes (Covaris). SMRT bell libraries were prepared according to the PacBio protocol recommendations. The libraries were purified with the 0.45x volume of AMPure PB beads and quantified on Qubit fluorometer using DNA HS kit. The insert size was determined on Bioanalyzer (Agilent) using DNA 12000 kit. SMRT bell libraries were pooled in the equimolar amount into a 20plex Microbial SMRT bell library according to the PacBio Microbial Multiplex Calculator. Conditioned sequencing primer 4 was annealed and Sequel II DNA Polymerase 2.0 was bound to the SMRT bell library. The complex was sequenced on PacBio Sequel II instrument, using 1 SMRT cell 8M and Sequencing chemistry 2.0, 15-hour sequencing movies were recorded.