GSM5733141: EtOH1; Mycobacterium tuberculosis CDC1551; RNA-Seq
Trizol LS (Thermo Fisher). Pellets were washed and suspended in Trizol LS (Ambion), 0.1 mmol silicon beads were added, and cells were lysed using a BeadBeater. Total RNA was isolated by chloroform extraction and precipitated in isopropanol with GlycoBlue reagent (Thermo Fisher). RNA was resuspended in nuclease-free water, genomic DNA contamination was removed using the Turbo-DNA free kit (Invitrogen), and rRNA was depleted using the Ribo-Zero Gold rRNA removal kit (Illumina). Sample quality was determined via Fragment Analyzer (Advanced Analytical) and TruSeq-barcoded RNAseq libraries were generated with the NEBNext Ultra II Directional RNA Library Prep Kit (New England Biolabs). Sequencing was performed at the Cornell University Transcriptional Regulation and Expression Facility on a NextSeq500 instrument (Illumina) at a depth of 15 M single-end 75 bp reads.