GSM8394236: OX_REP 3; Mycobacterium tuberculosis; RNA-Seq
5mL of bacterial culture were samples were pelleted, resuspended in TRIzol® Reagent (InvitrogenTM), and transferred to 2-mL screw cap tubes containing Zirconia/Silica beads. To achieve cellular disruption three rounds of 35-second pulses in a Mini Bead Beater (BioSpec Products), with each pulse alternated with 35-second incubations on ice were performed. Samples were finally centrifuged at maximum speed for 15 minutes at 13200 rpm at 4°C and Direct-zol RNA MicroPrep kit was used for RNA extraction. Qiagen QIAseq FastSelect – 5S/16S/23S was employed to perform rRNA depletion and stranded libraries were prepared using Illumina Stranded mRNA Prep.