title: PacBio CCS reads of whole genome sequencing of Bacillus thuringiensis strain BLB1
design: Single-molecule Real-time long reads sequencing was performed at Gentyane Sequencing Platform (Clermont-Ferrand, France) with a PacBio Sequel II Sequencer (Pacific Biosciences, Menlo Park, CA, USA). The SMRTBell library was prepared using a SMRTbell prep kit3, following the procedure and checklist -preparing whole genome and metagenome libraries using SMRTbell prep kit 3.0 protocol. Genomic DNA (10 ug) of each strain was sheared using g-tubes (Covaris, England) generating DNA fragments of approximately 10 kb. A Fragment Analyzer (Agilent Technologies, Santa Clara, CA, USA) assay was used to assess the fragments size distribution. Sheared genomic DNA were carried into the enzymatic reactions to remove the single-strand overhangs and to repair any damage that may be present on the DNA backbone. A A-tailing reaction followed by the barcoded overhang adapter ligation was conducted to generate the SMRT Bell templates. After nuclease treatment, the sample was then size-selected for fragments above 5kb with 35% AMPure PB Beads and equimolar multipled to obtain the final libraries around 10 kb. The SMRTBell libraries were quality inspected and quantified on a Fragment Analyzer (Agilent Technologies) and a Qubit fluorimeter with Qubit dsDNA HS reagent Assay kit (Life Technologies). A ready-to-sequence SMRTBell Polymerase Complex was created using a Binding Kit 3.2 (PacBio) and the Sequel II primer 3.2. The PacBio Sequel instrument was programmed to load a 90 pM library and sequenced in CCS mode on a PacBio SMRTcell 8M, with the Sequencing Plate 2.0 (Pacific Biosciences), 2 hours of pre-extension time and  acquiring one movie of 15 hours per SMRTcell.