GSM3937518: Pleura 1; Streptococcus pneumoniae; Mus musculus; RNA-Seq
Samples in Trizol with Zirconia beads from the animal model of infection were vortexed for 10 minutes to ensure complete lysis. Bacteria were also grown to mid-log phase in broth and processed in identical fashion. 1.33 ml of chloroform was added to each tube before vigorous shaking and incubation for 10 minutes at room temperature. Samples were then centrifuged at 8,000 g for 15 minutes at 4°C and the aqueous phase removed with a Pasteur pipette. RNA was extracted using the Zymo RNA clean and concentrator kit (Zymo Research) according to the manufacturer's instructions. Further purification was performed with Agencourt RNAClean XP beads (Beckman Coulter). Ribosomal RNA was depleted using the Ribo-Zero rRNA removal kit (Illumina) according to the manufacturer's instructions. RNA concentration and quality were assessed using the Agilent 2100 Bioanalyser. Strand specific ScriptSeq library prep kit (Illumina)
GEO Accession
GSM3937518