GSM5671207: HM7_iron_limited_rep3; Escherichia coli; RNA-Seq
GSM5671207
bacterial pellets were treated with lysozyme (Sigma, final concertation: 1 mg/mL) and proteinaseK (Qiagen, final concertation: 1.81 μg/mL) in RNAse-free Tris-EDTA buffer (Corning), for five minutes with continual vortexing. Total RNA was extracted using RNeasy mini kit (Qiagen). Genomic DNA was removed with Turbo DNA-free kit (Invitrogen). Bacterial rRNA was removed with the RiboMinus Transcriptome Isolation Kit, Bacteria (Invitrogen). rRNA was depleted using ThermoFisher's Ribominus Bacteria 2.0 Transcriptome Isolation kit. Libraries were prepared using NEBNext Ultra II Directional RNA Library Prep Kit as per manufacter's instructions