characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2014-05-30T17:02:17Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2013-11-22T17:15:37Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA2250796
characteristics.characteristic: External Id
characteristics.text: Ghent University
characteristics.characteristic: INSDC center name
characteristics.text: 2014-05-30T17:02:17Z
characteristics.characteristic: INSDC first public
characteristics.text: 2013-11-22T17:15:37Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS371962
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-2079:mut2
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: Protocols: an inoculum suspension containing approximately 5 x 107 CFU/ml was added to the wells of a round-bottomed 96-well microtiter plate (TPP). Ten wells were included per strain and the experiment was repeated twice (n = 3 x 10). Biofilms were grown in LBB supplemented with Tp and 0.2 % rhamnose to induce toxin expression. Following 4h of adhesion, the supernatant was removed and the plates were rinsed with physiological saline (PS). Subsequently, 100 ul of fresh LBB supplemented with Tp and 0.2 % rhamnose was added and the plates were further incubated at 37degreesC. after 24 h of growth, cells were harvested by vortexing (5 min) and sonication (5 min) and transferred to sterile tubes. RNA was extracted immediately following harvesting of the cells using the Ambion RiboPure Bacteria Kit (Life Technologies, Renfrewshire, UK), according to the manufacturerﾒs instructions. This procedure included a DNase I treatment for 1 h at 37degreesC. RNA quantifications were performed using picogreen fluorescence measurement on a FLUOstar galaxy plate reader (BMG Labtech, Ortenberg, Germany). Two microgram total RNA of each sample was depleted for ribosomal RNA using the Ribo-zero Magnetic Gram-Negative Bacteria kit (Epicentre, Madison, WI, U.S.A.). Approximately 30 ng of rRNA depleted RNA was used to create barcoded strand specific cDNA sequencing libraries with the Truseq stranded Total RNA library preparation kit (Illumina, San Diego, USA).
characteristics.characteristic: description
characteristics.text: overexpression of BCAM0258
characteristics.characteristic: genetic modification
characteristics.text: BCAM0258 overexpression mutant
characteristics.characteristic: genotype
characteristics.text: Burkholderia cenocepacia
characteristics.characteristic: organism
characteristics.text: Burkholderia cenocepacia
characteristics.characteristic: organism
characteristics.text: Burkholderia cenocepacia
characteristics.characteristic: scientific_name
characteristics.text: frozen specimen
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/OBI_0000922
characteristics.characteristic: specimen with known storage state
characteristics.text: J2315 or LMG16656
characteristics.characteristic: strain
characteristics.text: mut2
characteristics.characteristic: title