Wild-type Campylobacter jejuni NCTC11168 under iron replete conditions
C. jejuni NCTC11168 was routinely cultured on Mueller-Hinton (MH) agar plates under microaerophilic conditions (83% N2, 4% H2, 8% O2 and 5% CO2) at 37°C in a MACS-VA500 workstation. C. jejuni strains were grown overnight in MH agar overlaid with MH broth. Prior to inoculation, overnight cultures were washed in MEMα to remove excess iron sources. Glass flasks used for growing strains for RNA extraction were soaked with 1M HCl to solubilize bound metal ions and rinsed twice with ddH2O prior to sterilization. Strains were grown in a final volume of 50 mL MEMα supplemented with 10 mM pyruvate with freshly prepared FeSO4 added at a final concentration of 40 µM as needed to generate iron replete growth conditions. Library construction, cluster generation and sequencing were performed at the McGill University and Génome Québec Innovation. Ribosomal RNAs were depleted using the RiboZero Meta-Bacteria kit and strand-specific sequencing libraries were constructed using TruSeq kits. Cluster generation was done using standard Illumina protocols with sequencing performed on an Illumina HiSeq 2500 sequencer. The libraries were subjected to paired-end sequencing with a target read length of 100 nucleotides.