characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2016-03-13T17:07:02Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2016-03-03T14:42:36Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA3886794
characteristics.characteristic: External Id
characteristics.text: Microbial Genomics and Biotechnology Center for Biotechnology - CeBiTec Bielefeld University
characteristics.characteristic: INSDC center name
characteristics.text: 2016-03-13T17:07:02Z
characteristics.characteristic: INSDC first public
characteristics.text: 2016-03-03T14:42:36Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS1073928
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-4522:M_smegmatis_CO1
characteristics.characteristic: Submitter Id
characteristics.text: 1
characteristics.characteristic: biological replicate
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: Protocols: For stress experiments Mycobacterium smegmatis mc2155 wild type strain was grown in modified Hartmans-de Bont minimal medium (HdB) (finally per liter: EDTA [0.01 g], MgCl2x6H2O [0.1 g], CaCl2x2H2O [1 mg], NaMoO4x2H2O [0.2 mg], CoCl2x6H2O [0.4 mg] , MnCl2x2H2O [1 mg] , ZnSO4x7H2O [2 mg] , FeSO4x7H2O [5 mg] , CuSO4x5H2O [0.2 mg]; complemented with 15 mM (NH4)2SO4, 0.05% Tween80, 27.4 mM glycerol, 50 mM MOPS; 8.90 mM K2HPO4 and 7.08 mM NaH2PO4; pH 7.0) at 37°C and vigorous agitation. 15% sodium hypochlorite (NaOCl) was purchased from Sigma Aldrich. After harvesting control samples cultures were exposed to 1 mM NaOCl at an optical density at 500 nm (OD500) of 0.4 - 0.45 in the early log phase. Briefly, M. smegmatis mc2155 cells of 3 replicate experiments were harvested before and 30 min after exposure to 1 mM NaOCl and disrupted in 3 mM EDTA/ 200 mM NaCl lysis buffer with a Precellys24 Ribolyzer. RNA isolation was performed using phenol-chloroform-isoamylalcohol approach. After precipitation with 3 M sodium acetate and isopropanol, the total RNA was washed with cold ethanol and the pellet was solved in sterile water. Initially RNA quality was checked by Trinean Xpose (Gentbrugge,Belgium) and Agilent RNA Nano 6000 kit on Agilent 2100 Bioanalyzer (Agilent Technologies, Boblingen, Germany). Samples contaminated with DNA were treated with DNase (Qiagen), cleaned as described above and rechecked by Xpose and Agilent Bioanalyzer. Ribo-Zero rRNA Removal Kit (Bacteria) from Illumina (San Diego, CA, USA) was used to remove the ribosomal RNA molecules from the isolated total RNA. Removal of rRNA was checked by Agilent RNA Pico 6000 kit on Agilent 2100 Bioanalyzer (Agilent Technologies, Boblingen, Germany).
characteristics.characteristic: description
characteristics.text: Mycobacterium smegmatis str. MC2 155
characteristics.characteristic: organism
characteristics.text: Mycobacterium smegmatis str. MC2 155
characteristics.characteristic: organism
characteristics.text: Mycobacterium smegmatis str. MC2 155
characteristics.characteristic: scientific_name
characteristics.text: M_smegmatis_CO1
characteristics.characteristic: title