characteristics.text: 2013-06-12T17:02:04Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2013-01-21T13:01:28Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA1920909
characteristics.characteristic: External Id
characteristics.text: Imperial College
characteristics.characteristic: INSDC center name
characteristics.text: 2013-06-12T17:02:04Z
characteristics.characteristic: INSDC first public
characteristics.text: 2013-01-21T13:01:28Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS207721
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-1456:Cell culture 4
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: Protocols: M. smegmatis mc2 155 wild type (ATCC 700084) and M. smegmatis mc2 155 _glnR strains were used in this study. M. smegmatis was grown aerobically in Middlebrook 7H9 liquid broth (supplemented with 0.2% glycerol, 0.05% Tween 80 and 10% OADC) at 37 C, 180 rpm for 24 hours. For growth in nitrogen limiting conditions, 5 ml culture of M. smegmatis was washed twice in nitrogen free Sauton's medium (0.05% (w/v) KH2PO4, 0.05% (w/v) MgSO4, 0.2% (w/v) citric acid, 0.005% (w/v) ferric citrate, 0.2% (v/v) glycerol, 0.0001% (v/v) ZnSO4, 0.015% (v/v) Tyloxapol) and inoculated into 30 ml Sauton's nitrogen free medium, supplemented with 30mM (nitrogen non-limiting) ammonium sulphate to a starting OD600 of 0.08. Cultures were grown at 180 rpm 37 C. Growth was monitored by OD600. Ammonium ions in the culture medium were quantified using an AquaQuant Ammonium detection kit (Merck). M. smegmatis strains were grown in triplicate in nitrogen limiting conditions as stated until external nitrogen had completely depleted. Cells were immediately added to final concentration 2.5 M GTC and harvested by centrifugation. Pellets were resuspended in 1 ml TRIzol (Life Technologies) and stored at -80 C. Thawed cell/Trizol suspensions were transferred to tubes containing Zirconium beads (MP Biomedicals) and the cells lysed in a ribolyser Fastprep (Hybaid). RNA samples were chloroform extracted and RNA purified using the RNeasy kit (Qiagen) according to manufacturer's instructions.
characteristics.characteristic: description
characteristics.text: Mycobacterium smegmatis
characteristics.characteristic: organism
characteristics.text: Mycobacterium smegmatis
characteristics.characteristic: organism
characteristics.text: wild type
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0005168
characteristics.characteristic: phenotype
characteristics.text: Mycobacterium smegmatis
characteristics.characteristic: scientific_name
characteristics.text: mc2 155 (ATCC 700084)
characteristics.characteristic: strain or line
characteristics.text: 13 hour
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/UO_0000032
characteristics.characteristic: time
characteristics.text: Cell culture 4
characteristics.characteristic: title