characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2014-10-25T17:01:04Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2018-03-08T21:08:54Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA2765963
characteristics.characteristic: External Id
characteristics.text: DKFZ/ZMBH Alliance, University of Heidelberg, Heidelberg, Germany
characteristics.characteristic: INSDC center name
characteristics.text: 2014-10-25T17:01:04Z
characteristics.characteristic: INSDC first public
characteristics.text: 2018-03-08T21:08:54Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS542740
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-2915:UAS-esg no4
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: fruit fly
characteristics.characteristic: common name
characteristics.text: esgts > UASt-esg Protocols: esg-Gal4ts flies were crossed to either w1118 (esgtsCon), UASt-esg (UAS-esg) or esgRNAiJF03134 (esgRNAi) and shifted for 24 hours to 29 degrees C to induce transgene expression. 80-90 midguts were prepared for each genotype. For midgut cell dissociation for RNA-Seq, 1 mg/ml of Elastase were added per sample and guts incubated for 1 h at 27 degrees C, shaking at 600 rpm. To help dissociation, samples were pipetted vigorously every 15 min. For FACS profiling, tissue dissociation was performed by treatment with 7.5 mg/ml of Collagenase for 45 min followed by addition of 50 ul of 10X Trypsin/EDTA (Sigma) for 15-20 min. For RNA-Seq, a minimum of 20,000 GFP+ cells/condition was sorted on a FACS Aria II Flow Cytometer (BD Biosciences) using a 70 um nozzle and FACS Diva software. Intestinal cells obtained from w1118 animals where used to control for auto-fluorescence and set sorting gates. Dead cells were excluded from the sorting procedure by staining with 3 ul of Propidium Iodide (1 mg/ml, Life Technologies/Molecular Probes). RNA was isolated from sorted cells using the Arcturus PicoPure RNA Isolation Kit (Life Technologies). mRNA was amplified using the Arcturus RiboAmp HS Plus Amplification Kit (Life Technologies). Amplified RNA integrity was determined with an Agilent 2100 Bioanalyzer before library generation. cDNA libraries were generated using the NEBNext Ultra RNA Library Prep Kit for Illumina (NEB), and NEBNext Multiplex Oligos for Illumina (NEB) were used for multiplexing.
characteristics.characteristic: description
characteristics.text: UAS-esg overexpression
characteristics.characteristic: genotype
characteristics.text: Drosophila melanogaster
characteristics.characteristic: organism
characteristics.text: Drosophila melanogaster
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_7227
characteristics.characteristic: organism
characteristics.text: Drosophila melanogaster
characteristics.characteristic: scientific_name
characteristics.text: UAS-esg no4
characteristics.characteristic: title