title: Whole genome sequencing of Bacillus thuringiensis
design: Bacillus thuringiensis strains were revived from glycerol stocks by streaking onto Luria-Bertani (LB) agar plates. A single colony was inoculated into 5 mL LB broth and cultured overnight at 37C . Subsequently, 0.3 mL of the activated culture was transferred into 30 mL LB broth and incubated for ~3 hours until turbid. Cells were harvested by centrifugation and washed with Sodium chlorideTrisEDTA (STE) buffer. Genomic DNA was extracted following standard protocols. Briefly, cell pellets were treated with lysozyme (100 mg/mL) at 4C overnight, followed by Rnase A digestion and SDS-mediated lysis. Protein and impurities were removed using phenol:chloroform:isoamyl alcohol extraction, and DNA was precipitated with ethanol and sodium acetate. The DNA pellet was washed, air-dried, and resuspended in Tris-EDTA (TE) buffer. DNA quality was assessed by 0.8% agarose gel electrophoresis. High-quality genomic DNA samples of Bacillus thuringiensis isolates were subjected to whole-genome sequencing using the Illumina HiSeq 2500 platform.