Whole genome sequence of Bacillus subtilis
Bacteria were grown in Nutrient broth for 16 hours with 180 rpm at 34C. Genomic DNA was isolated following the method of Sambrook and Russel. The quantity and purity of DNA was assessed using a NanoDrop 2000 spectrophotometer (NanoDrop, Thermo Scientific). The integrity of DNA was evaluated by 1% agarose gel electrophoresis. Nextera XT libraries were prepared manually following the manufacturers protocol (Illumina). Briefly, samples were normalized to 2.5 ng/l DNA material per library using a Quant-iT picogreen assay system (Life Technologies) on an AF2200 plate reader (Eppendorf), then fragmented by enzymatic treatment and tagged via tagmentation. Amplification was executed by Veriti 96 well PCR (Applied Biosystems) trail around with AMPure XP bead cleanup (Beckman Coulter). Sequencing was performed on an Illumina NextSeq 500 sequencer.