characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2014-02-27T17:00:44Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2018-03-08T17:12:56Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA2377778
characteristics.characteristic: External Id
characteristics.text: European bioinformatic Institute
characteristics.characteristic: INSDC center name
characteristics.text: 2014-02-27T17:00:44Z
characteristics.characteristic: INSDC first public
characteristics.text: 2018-03-08T17:12:56Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS408383
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-1822:NPC_3.30
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: neuro progenitor cell
characteristics.characteristic: cell type
characteristics.text: house mouse
characteristics.characteristic: common name
characteristics.text: Protocols: ESCs were cultured using standard procedures in medium containing 1,000U/ml leukemia inhibitory factor (Millipore) with irradiated MEF feeders (Global Stem) on gelatin-coated plates. ESCs were removed from feeder cells by soaking twice on gelatin-coated plates for 1 hour each prior to sample collection or cell differentiation. Differentiation was performed by culturing ESCs in the absence of MEF feeders in 50:50 DMEM/F12:Neurobasal medium (Gibco) supplemented with 1x N2 (Gibco), 1x B27 (Gibco), 40mg/L insulin (Sigma), 25ug/ml BSA Fraction V (Gibco) at 0.5x106-2.0x106 cells/10cm plate for 6 days. Cells were then resuspended in N2 expansion medium (DMEM/F12, 50ug/ml BSA fraction V, 10ng/ml EGF (Preprotech), 10ng/ml FGF (Preprotech), 1ug/ml laminin (Invitrogen), 1x N2) and plated onto uncoated T75 flasks to allow for neurosphere outgrowth. Following 4 days, neurospheres were collected by mild centrifugation and plated onto gelatin-coated plates in N2 expansion medium. Following 2-3 passages, cells represented a homogeneous population of NPCs, which could be seeded to expand single-cell derived clones. RNA from was isolated using Trizol reagent (Ambion) according to the manufacturer's instructions. PolyA+ RNA was isolated (Oligotex kit, Qiagen) from 10ug total RNA. Stranded libraries were prepared using a protocol adapted from (Parkhomchuk et al., 2009) for paired-end sequencing on the Illumina GA IIx platform.
characteristics.characteristic: description
characteristics.text: Mus musculus
characteristics.characteristic: organism
characteristics.text: Mus musculus
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_10090
characteristics.characteristic: organism
characteristics.text: clonal expansion
characteristics.characteristic: passage
characteristics.text: Mus musculus
characteristics.characteristic: scientific_name
characteristics.text: fresh specimen
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/OBI_0000971
characteristics.characteristic: specimen with known storage state
characteristics.text: C57BL6xCAST
characteristics.characteristic: strain
characteristics.text: NPC_3.30
characteristics.characteristic: title