Streptococcus agalactiae isolates were cultured on blood agar plates at 30&deg;C, then transferred to tryptic soy broth with yeast extract and incubated until mid-logarithmic phase. Bacterial pellets were collected by centrifugation and stored at -80&deg;C. Genomic DNA was extracted using a modified Qiagen DNeasy Blood &amp; Tissue Kit protocol. DNA quantity and quality were assessed spectrophotometrically and by agarose gel electrophoresis. Sequencing library preparation was performed using QIASeq FX DNA Library Preparation protocol according to manufacturer&#39;s instructions. The amplified library was purified using Cambrian Bioworks CamSelect NGS followed by library quality control check. Illumina-compatible sequencing libraries were quantified by Qubit fluorometer and fragment size distribution was analyzed on an Agilent 4150 TapeStation. Illumina compatible libraries were converted to Onso compatible libraries by Onso Library Conversion PCR.