Cultivation of Livestock-Associated MRSA ST398

The whole genome sequenced wild type (WT) livestock-associated methicillin-resistant S. aureus ST398 (Genbank accession AM990992) [21] (link) and S. aureus RN4220 were grown in Brain Heart Infusion (BHI) (Oxoid, Difco) broth at 37°C with aeration. S. aureus SH1000 pMARGH2b, S. aureus SH1000 pFA545 and S. aureus RN4220 pFA545gen were grown in BHI or Tryptic Soy Broth (TSB) (Oxoid) with 5 mg/l erythromycin (Sigma), 5 mg/l tetracycline (Sigma) and 16 mg/l gentamicin (Sigma) respectively, at 30°C with aeration. For solid growth BHI agar, sheep blood agar plates (Oxoid) or Tryptic Soy Agar (TSA) (Oxoid) were applied and supplemented with the appropriate antibiotic if needed. Escherichia coli DH10 was cultured in Luria Broth (LB) at 37°C with aeration or on LB agar plates (Sigma).