characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2017-08-29T17:02:00Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2017-03-13T15:04:19Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA103910533
characteristics.characteristic: External Id
characteristics.text: London School of Hygiene and Tropical Medicine
characteristics.characteristic: INSDC center name
characteristics.text: 2017-08-29T17:02:00Z
characteristics.characteristic: INSDC first public
characteristics.text: 2017-03-13T15:04:19Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS1599701
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-5557:Sample 2
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: Protocols: Mycobacterium tuberculosis H37Rv (SysteMTb) was grown in Middlebrook 7H9 medium supplemented with 0.4% glycerol, 0.085% NaCl, 0.5% BSA and 0.05% Tyloxapol in roller bottle culture (2 rpm at 37°C). For nitric oxide experiments, cultures were grown until mid-exponential phase and Diethylenetriamine/nitric oxide adduct (deta/NO, Sigma) was added to a final concentration of 1 mM. For transcriptomic analyses, samples were harvested from triplicate cultures immediately before addition of nitric oxide and 20 minutes, 2 and 24 hours after nitric oxide addition. For each sample, 25 mL of culture were spun down and isolation of RNA was performed using the FastRNA Pro blue kit from QBiogene/MP Bio according to manufacturer’s instructions. Cultures were first rapidly cooled by addition of ice directly to the culture prior to centrifugation. All RNA samples were treated with Turbo DNase free (Ambion) until any DNA contamination removed. Concentration and quality control of RNA samples was measured by Nanodrop (ND-1000, Labtech) and Agilent RNA chip (2100 Bioanalyser). For construction of directional RNA-seq libraries, Ribo-Zero™ and ScriptSeq™ v2 Kits (Epicentre) were used. Briefly, 5 µg of total RNA were used for ribosomal RNA removal using the Ribo-Zero rRNA removal kit (Gram-Positive Bacteria) following manufacturer’s instructions. 50 ng of Ribo-Zero-treated RNA were used to construct cDNA libraries for RNA sequencing using the ScriptSeq™ v2 Kit.
characteristics.characteristic: description
characteristics.text: Mycobacterium tuberculosis H37Rv
characteristics.characteristic: organism
characteristics.text: Mycobacterium tuberculosis H37Rv
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_83332
characteristics.characteristic: organism
characteristics.text: Mycobacterium tuberculosis H37Rv
characteristics.characteristic: scientific_name
characteristics.text: Sample 2
characteristics.characteristic: title