RNA-Seq of Staphylococcus aureus
S. aureus strains grown overnight on blood agar plates (BAPs) (37C with 5% CO2) were diluted in tryptic soy broth (TSB) to achieve a turbidity equivalent of 4.0 McFarland standard. Following a 2 h incubation at 37C under stationary conditions, the cultures were centrifuged at 4,000 rpm for 15 min at 4C to pellet the cells. Sbsequently, 0.5 MIC (128 g/mL) thymol was added to the remaining culture broth and incubated under similar conditions at 37C. After a further 3 h incubation, the culture broth was collected and centrifuged to pellet the cells. RNA was extracted and purified using the RNeasy Mini Kit (Qiagen, Germany) according to the manufacturers protocol. RNA quality and quantity were analyzed using a NanoDrop 1000TM spectrophotometer (Thermo Fisher Scientific) and the 2100 Bioanalyzer from Agilent Technologies, respectively. The RNA-Sequencing was performed using the high-throughput Illumina HiSeq2500 sequencing platform.