GSM8330378: H37Rv WT Rep1; Mycobacterium tuberculosis H37Rv; RNA-Seq
Bacterial strains were pelleted and bead beated in 1 mL of TRIzol (Life Technologies) with 0.1 mm silica beads (MP Biomedicals). After centrifugation, supernatants were extracted with chloroform, and RNA was precipitated with isopropanol and glycogen. RNA pellets were washed with 75% ethanol and dissolved in RNase-free water. Contaminant DNA was removed by incubation with DNase (TURBO DNA-free kit, Life Technologies). RNA cleanup was performed with the RNeasy Mini Kit (Qiagen). RNA integrity and quality were assessed with an Agilent Bioanalyzer device (Agilent Technologies). RNA-seq libraries were prepared using the Stranded Total RNA Prep and Ligation with Ribo-Zero Plus kit (Illumina) and sequenced using a NextSeq 2000 device (Illumina).