GSM4279912: JE84-2-15; Streptococcus pyogenes; RNA-Seq
In vitro grown pelleted cells were mechanically disrupted using silica beads, RNA was isolated using RNeasy kit, contaminating DNA was degraded using Turbo DNA-free (Ambion), and rRNA was depleted using Ribo-Zero (Illumina). The tissue (80-100 mg) corresponding to the in vivo samples was washed in PBS, homogenized, and mechanically disrupted using 0.1 mm and 0.5 mm beads. RNA was extracted (MasterPure Complete DNA and RNA purification kit, Lucigen), subjected to three consecutive DNase treatments with Turbo DNA-free (Ambion), and depletion of ribosomal RNA was performed with the Ribo-Zero Gold rRNA Removal Kit, Epidemiology (Illumina). Resultant RNA from dahA mutant and parental wild-type samples was reverse transcribed to cDNA and sequencing libraries were generated using ScriptSeq RNA-seq preparation kit (Epicentre). cDNA libraries from in vivo samples were generated from rRNA depleted RNA using a NEBNext Ultra II Directional RNA Library Preparation kit for Illumina (NEB).
GEO Accession
GSM4279912