title: NextSeq 500 paired end sequencing; RNA-seq of coding RNA of Staphylococcus aureus COL wild type strain before and after 30 min of exposure to 176 μM neutrophil-derived oxidant hypothiocyanous acid (HOSCN) during exponential phase of an OD540 of 0.5 in LB medium
design: RNA-seq of coding RNA of Staphylococcus aureus COL wild type strain before and after 30 min of exposure to 176 μM neutrophil-derived oxidant hypothiocyanous acid (HOSCN) during exponential phase of an OD540 of 0.5 in LB medium
library_construction_protocol: Cells were cultivated in LB medium in 3 biological replicates and harvested at an OD500 of 0.5 before (as control) and at 30 min after exposure to 176 μM neutrophil-derived oxidant hypothiocyanous acid (HOSCN) stress. Bacteria were harvested fast centrifugation. The pellets were immediately frozen in liquid nitrogen and stored at -80°C. Staphylococcus aureus COL  was cultivated in LB medium in 3 biological replicates and harvested at an OD500 of 0.5 before (as control) and at 30 min after exposure to 1176 μM neutrophil-derived oxidant hypothiocyanous acid (HOSCN) stress Staphylococcus aureus COL  was cultivated in LB medium in 3 biological replicates and harvested at an OD500 of 0.5 without any treatment Cells were disrupted in 3 mM EDTA/ 200 mM NaCl lysis buffer with a Precellys24 ribolyzer followed by RNA isolation using the acid phenol extraction protocol. The RNA quality was approved by Trinean Xpose (Gentbrugge, Belgium) and the Agilent RNA Nano 6000 kit using an Agilent 2100 Bioanalyzer (Agilent Technologies, Böblingen, Germany). Ribo-Zero rRNA Removal Kit (Bacteria) from Illumina (San Diego, CA, USA) was used to remove the rRNA. TruSeq Stranded mRNA Library Prep Kit from Illumina (San Diego, CA, USA) was applied to prepare the cDNA libraries.