Whole-genome sequencing of Campylobacter ureolyticus
Archived isolates were revived and maintained on BD BBL trypticase soy agar with 5% sheep blood (Blood agar plate, BD) at 35C under anaerobic conditions (5% CO2, 5% H2, 90% N2) in an AS-580 Anaerobe Chamber (Anaerobe Systems). Growth was resuspended in 1 mL of Tryptic Soy Broth (TSB) and cells were pelleted by centrifugation at >5000g for 10 minutes. Following removal of supernatant, genomic DNA was extracted with the DNeasy Blood & Tissue Kit according to the manufacturers instructions for Gram-negative Bacteria (Qiagen). Extracted DNA was quantitated using the QuantiFluor dsDNA System (Promega) and a DS-11 Spectrophotometer/Fluorometer (DeNovix). Sequencing libraries were prepared using a DNA Prep Kit (Illumina) according to the manufacturers instructions and quantitated with the QuantiFluor dsDNA System. Paired-end sequencing was performed with a 500-cycle MiSeq Reagent Kit v2 on a MiSeq instrument (Illumina).