Illumina HiSeq 1000 paired end sequencing; RNA-seq of coding RNA of Staphylococcus aureus USA300 wild type strain before and after 30 min of exposure to 1.5 mM in RPMI HOCl stress during the log phase in RPMI media
RNA-seq of coding RNA of Staphylococcus aureus USA300 wild type strain before and after 30 min of exposure to 1.5 mM in RPMI HOCl stress during the log phase in RPMI media
Cells were cultivated in RPMI medium in 3 biological replicates and harvested at an OD500 of 0.5 before (as control) and at 30 min after exposure to 1.5 mM HOCl stress. Bacteria were harvested fast centrifugation. The pellets were immediately frozen in liquid nitrogen and stored at -80°C. Staphylococcus aureus USA300 was cultivated in RPMI medium in 3 biological replicates and harvested at an OD500 of 0.5 Staphylococcus aureus USA300 was cultivated in RPMI 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.