title: Illumina HiSeq 1000 paired end sequencing; RNA-seq of coding RNA of Streptococcus aureus wild type strain before and after 30 min of exposure to 13 mM itaconic acid stress reveals adaption to the immunometabolite itaconate by inducing acid and oxidative stress responses
design: RNA-seq of coding RNA of Streptococcus aureus wild type strain before and after 30 min of exposure to 13 mM itaconic acid stress reveals adaption to the immunometabolite itaconate by inducing acid and oxidative stress responses
library_construction_protocol: 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 13 mM itaconic acid stress. Bacteria were harvested fast centrifugation. The pellets were immediately frozen in liquid nitrogen and stored at -80°C. S. aureus COL was cultivated in RPMI medium in 3 biological replicates and harvested at an OD500 of 0.5  S. aureus COL was cultivated in RPMI medium in 3 biological replicates and was exposed/treated 30 min with 13 mM itaconic acid stress at an OD500 of 0.5  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.