Whole genome sequence of Bacillus licheniformis strain B65 recovered from the surface of a dairy pasteurizer after cleaning and disinfection

Whole genome sequence of Bacillus licheniformis strain B65 recovered from the surface of a dairy pasteurizer following cleaning and disinfection in the FASTA format. The bacteria was grown overnight in BHI broth and its DNA was extracted using the Qiagen tissue culture DNA extraction kit (DNeasy Blood & Tissue Kits). The genomic DNA was randomly sheared into short fragments. The obtained fragments were end repaired, A-tailed and further ligated with Illumina adapter. The library was checked with Qubit and real-time PCR for quantification and bioanalyzer for size distribution detection. Quantified libraries were pooled and sequenced on Illumina platform (NovaSeq 6000) according to effective library concentration and data amount required. The genome assembly process for sequence submission involved several meticulous steps using Clean Data. Initially, SOAPdenovo software was employed, where different K-mers (typically 95, 107, 119) were selected based on the project type. The optimal K-mer was determined, and other parameters were adjusted to choose the preliminary assembly with the fewest scaffolds. Next, SPAdes software was used with default K-mers of 99 and 127 to achieve an assembly with minimal scaffolds. Following this, Abyss software was utilized with a K-mer of 64 for assembly. The outcomes from these three software tools were then integrated using CISA software, selecting the result with the least scaffolds. GapClose software played a crucial role in filling gaps in the preliminary assembly. Additionally, contamination was minimized by removing reads with a sequencing depth lower than 0.35 times the average depth. Fragments shorter than 500 bp were filtered out, and the final assembly was prepared for gene prediction after these rigorous steps. Overall genome coverage was over 99%.