Bacillus subtilis NRS7692 Genomic Sequencing
Genomic DNA was extracted using the QIAprep Spin Miniprep Kit (cat. Nos. 27104 and 27106) and following the 'Quick-Start Protocol'. DNA concentration was measured using a Qubit 4 Fluorometer or a NanoDrop One/OneC Microvolume UV-Vis Spectrophotometer. NEBNext Ultra II DNA Library Preparation kit for was used for DNA library preparation following the manufacturer�s recommendations (NEB, Ipswich, MA, USA). Briefly, genomic DNA was fragmented by acoustic shearing with a Covaris instrument. Fragmented DNA was end repaired and adenylated. Adapters were ligated after adenylation of the 3�ends followed by enrichment by limited cycle PCR. Adapter-ligated DNA libraries were cleaned up and validated using Agilent TapeStation and quantified using Qubit 4.0 Fluorometer. The libraries have been multiplexed on a flowcell and loaded on the Illumina NovaSeq 6000 instrument according to the manufacturer's instructions. The samples were sequenced using a 2x150 paired-end (PE) configuration. Image analysis and base calling was conducted by the NovaSeq Control Software on the NovaSeq instrument. Raw sequence data (.bcl files) generated from Illumina NovaSeq was converted into fastq files and de-multiplexed using Illumina bcl2fastq program version 2.20. One mismatch was allowed for index sequence identification.