characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2014-10-07T17:00:59Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2018-03-08T19:38:08Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA2629794
characteristics.characteristic: External Id
characteristics.text: EBI
characteristics.characteristic: INSDC center name
characteristics.text: 2014-10-07T17:00:59Z
characteristics.characteristic: INSDC first public
characteristics.text: 2018-03-08T19:38:08Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS499327
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-2316_newruns:S2-RNAseq-groGFP-rep2
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: S2R+
characteristics.characteristic: cell line
characteristics.text: fruit fly
characteristics.characteristic: common name
characteristics.text: Protocols: Kc167 and S2R+ cells (obtained from the Drosophila Genomics Resource Centre [DGRC]) were grown in Schneiders Insect Cell Medium (GIBCO) supplemented with 10% FBS (Sigma) and 1% penicillin/streptomycin (Thermoscientific). Cell lines were kept at 25C. For Groucho oligomerization experiments cells were cotransfected with dsRNA against the 3UTR of Gro as well as plasmids containing either Gro-GFP or GroL38D;L87D-GFP. Transfections were performed using Effectene according to manufacturers instructions (Qiagen). 300-600ng DNA and 1 microgram of dsRNA were cotransfected per 1-3 x 10^6 cells. Successfully transfected cells were processed six days post transfection. For RNA-seq experiments total RNA was obtained from approximately 5 x 10^6 cells per sample using the Qiagen RNeasy mini kit. mRNA was then extracted using the Dynabeads mRNA Purification Kit (Life Technologies). 50-250 ng of mRNA was used to generate an mRNA library following manufacturers instructions (NEBnext mRNA Library Prep Master Mix _ E6110S). cDNA fragments of 250-350 bp were selected by agarose gel extraction and amplified with 12 cycles of PCR. Resulting libraries were assessed for concentration and size using the Qubit fluorometer and the Agilent2100 Bioanalyzer.
characteristics.characteristic: description
characteristics.text: gro-GFP and gro RNA
characteristics.characteristic: genetic modification
characteristics.text: gro-GFP and gro RNA
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: S2-RNAseq-groGFP-rep2
characteristics.characteristic: title