title: rnaseq_rhoDUC_0h_r1
organism: Mycobacterium tuberculosis H37Rv
description: Protocols: Mycobacterium tuberculosis H37Rv and RhoDUC were cultured in liquid Middlebrook 7H9 supplemented with 10% ADC (Sigma), 0.5% Glycerol and 50 µg/ml hygromycin if appropriate and 0.05% Tween 80. Cultures were harvested at an OD600nm ~ 0.6 for exponential phase. Cells were stopped using 30% ice, centrifuged at 5000rpm for 10min at 4°C and the RNA extracted using the FastRNA Pro Blue Kit (MP Biomedicals) following the instruction of the supplier. RNA concentration and purity was assessed using Nanodrop 2000 (ThermoFisher). RNA integrity was assessed using 2100 Bioanalyzer (Agilent) for RNA sequencing. DNA-free total RNA were fragmented with ultrasounds 4 pulses of 30 sec at 4°C and the library constructed with the TruSeq RNA library Kit V2 (Illumina).