RNA sequencing of Mycobacterium tuberculosis: isolate SAWC 5527 Mid log phase growth
Cultures were grown in Middlebrook 7H9 liquid media with the addition of dextrose and catalase, with growth phases determined by monitoring optical density over time. Samples for RNAseq were extracted during early logarithmic phase (Elog), middle logarithmic phase (ML(c)), and stationary phase (Stat). A fourth set of samples was treated during ML(C)) with hydrogen peroxide (H2O2) to simulate growth in the phagosomal environment (ML(T)). Each condition had 3 biological replicates, resulting in 24 total samples sequenced. For the removal of rRNA from the total RNA samples the Truseq stranded mRNA library preparation kit (RS-122-2101) with the Bacterial Ribozero kit (MRZMB126) was used. The samples were sequenced on an Illumina HiSeq 2500 using version 4 SBS chemistry (2x125bp), with approximately 10 million reads per sample loaded into the lane.
SAWC-5527-MLC-2