GSM6383311: S. aureus 3xMIC 3; Staphylococcus aureus; RNA-Seq
For total RNA extraction, 1 mL culture was harvested (5 min, 8,000  ´ g, RT), the cells were resuspended in 1 mL RNA protect buffer (New England Biolabs, Frankfurt am Main, Germany) and incubated for 5 min at RT. The cells were resuspended in 200 µL TE buffer with 25 µg lysostaphin and cell lysis was achieved by incubation at 37 °C for 30 min. RNA was extracted using the Monarch™ Total RNA Miniprep Kit (New England Biolabs) following the manufacturer's instructions and stored at −80 °C, before submission to the Genewiz sequencing facility (Leipzig, Germany). RNA samples were quantified using Qubit 4.0 Fluorometer (Life Technologies, Carlsbad, CA, USA) and RNA integrity was checked with RNA Kit on Agilent 5300 Fragment Analyzer (Agilent Technologies, Palo Alto, CA, USA). rRNA depletion was performed using NEBNext rRNA Depletion Kit (Bacteria). RNA sequencing library preparation was performed using NEBNext Ultra II RNA Library Prep Kit for Illumina by following the manufacturer's recommendations (NEB, Ipswich, MA, USA).