GSM3860726: 7H9_OADC3; Mycobacterium tuberculosis; RNA-Seq
Bacterial RNA extraction was performed using the protocol previously described (Rohde et al., 2012). See manuscript for reference. rRNA removal was performed using 50-100ng total RNA input and a modified protocol for the  Ribo-Zero Epidemiology Gold rRNA removal kit (Illumina). Briefly, 90 μl bead stock was used  per sample, together with 2 μl each of reaction buffer and removal solution in a 20 μl reaction  volume, as detailed in the manufacturer's protocol. The rRNA-depleted samples were purified by  precipitating the RNA. Sequencing libraries were generated using the NEBNext Ultra™ II  Directional RNA Library Prep Kit for Illumina (New England BioLabs). Libraries were sequenced  on a NextSeq500 (Illumina) in multiple rounds until the desired sequencing depth for bacterial  reads was reached (target 1M 85nt reads). Rna-seq Directional Libraries