GSM8046228: HG001 (p)ppGpp0, replicate 1; Staphylococcus aureus; RNA-Seq
Bacteria were harvested by centrifugation. Bacterial pellets were immediately resuspended in TRIzol (Thermo Fisher Scientific) and mechanically lysed using zirconia/silica beads in a high speed homogenizer. RNA was further purified using the ExpressArt® RNA ready Add-on Kit for TRIzol extraction (AmpTec). DNase digest of the sample was directly performed on the column. Per sample, a total amount of 100 ng RNA was subjected to rRNA depletion followed by cDNA library construction with the IlluminaTM Stranded Total RNA Prep Ligation with Ribo Zero Plus Kit according to the manufacturer's instructions. Libraries were sequenced as single-read (100 bp read length) on a NextSeq platform (Illumina) using the NextSeqTM 500 Mid Output Kit v2.5.