GSM7498445: Mt03_Sauton_rep4; Mycobacterium tuberculosis; RNA-Seq
RNA was isolated from exponentially growing cultures equivalent to 10 O.D.600units. The collected cells were resuspended in TRIzol reagent (Invitrogen), lysed using a bead beater (MP FastPrep system, MP Biomedicals) as per manufacturer's instructions, and the TRIzol-lysed cell mix was extracted with chloroform, followed by precipitation of RNA using isopropanol. The RNA pellet was dissolved in RNase-free water (30µl with 2µl of RiboLock (Thermo Scientific). DNA contamination was removed by DNase I (Invitrogen) treatment, and the RNA was further purified using RNeasy mini columns (Qiagen). Total RNA was quantified using Qubit RNA BR kit and quality was assessed using Agilent 4200 Tapestation. Libraries were generated using Illumina stranded total RNA ribozero plus. Final library was quantified using Qubit ds DNA HS kit and sequenced on Illumina NovaSeq 6000 on SP flow cell.