characteristics.text: 2013-11-25T17:03:13Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2013-04-17T14:30:30Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA2064879
characteristics.characteristic: External Id
characteristics.text: MRC National Institute for Medical Research
characteristics.characteristic: INSDC center name
characteristics.text: 2013-11-25T17:03:13Z
characteristics.characteristic: INSDC first public
characteristics.text: 2013-04-17T14:30:30Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS237630
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-1616:Starv_R3_WT
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 starvation experiments, exponentially growing bacteria were washed, resuspended in PBS supplemented with 0.025% Tyloxapol, and maintained in roller bottle culture for a further 24 hours RNA was isolated from triplicate exponentially growing and PBS-washed cultures using the FastRNA Pro blue kit from QBiogene/MP Bio according to manufactures instructions. All RNA samples were treated with Turbo DNase free (Ambion). For the synthesis of whole transcriptome cDNA, RNA was fragmented with ultrasound (4 pulses of 30 s at 4oC), then treated with Antarctic phosphatase and re-phosphorylated with polynucleotide kinase (PNK). Fragmented RNA was then poly(A)-tailed using poly(A) polymerase and a RNA adapter was ligated to the 5'-phosphate of the RNA. First-strand cDNA synthesis was performed using an oligo(dT)-adapter primer and M-MLV reverse transcriptase. The resulting cDNA was PCR-amplified to about 20-30 ng/?l using a high fidelity DNA polymerase. For TSS mapping, RNA was enriched for primary transcripts by incubating fragmented, PNK-treated samples with Terminator exonuclease (TEX, Epicentre) which specifically degrades RNA species carrying a 5' monophosphate (5'P). Exonuclease-resistant RNA species (primary transcripts with 5'PPP) were poly(A)-tailed using poly(A) polymerase, followed by treatment with tobacco acid pyrophosphatase (TAP, Epicentre) to degrade 5'PPP to 5'P RNA
characteristics.characteristic: description
characteristics.text: Starved cultures
characteristics.characteristic: growth condition
characteristics.text: Mycobacterium tuberculosis
characteristics.characteristic: organism
characteristics.text: Mycobacterium tuberculosis
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_1773
characteristics.characteristic: organism
characteristics.text: Mycobacterium tuberculosis
characteristics.characteristic: scientific_name
characteristics.text: H37Rv
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_83332
characteristics.characteristic: strain
characteristics.text: 24 hour
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/UO_0000032
characteristics.characteristic: time
characteristics.text: Starv_R3_WT
characteristics.characteristic: title