organism: Homo sapiens
description: Protocols: RNA was isolated from 12 NPC lines (8 control and 4 case lines). All iPSCs were derived from human donor skin fibroblasts using integration-free episomal methods. Plasmids incorporating Oct4/shP53, SOX2/Klf4 and L-Myc/Lin28 (pCXLE-hSK and pCXLE-hUL, Addgene plasmids 27077, 27078, 27080) were electroporated into dermal fibroblasts using an Amaxa Nucleofection Kit (Lonza). Colonies were selected between day 21-25 and individual colonies of cells were expanded into single wells coated with Geltrex (Life Technologies) in Essential 8 media (Life Technologies) at 37C and 5% CO2. Prior to neuralisation iPSC were transitioned to reduced-growth factor Matrigel (BD Biosciences) coated plastic dishes. Anterior NPCs were then derived from parental pluripotent stem cell lines. iPS cells were neurally converted in suspension in CDM as described in Bilican et al. \Physiological Normoxia and Absence of EGF Is Required for the Long-Term Propagation of Anterior Neural Precursors from Human Pluripotent Cells\, PLoS One (2014), 9(1): e85932. RNA was extracted from cell pellets using the RNeasy Mini kit (QIAGEN). Genomic DNA was removed with the DNA-Free kit (Ambion). QC analysis using an Agilent 2100 bioanalyzer demonstrated that all samples had RNA integrity number (RIN) values between 7.7-10. Libraries were prepared with the TruSeq Stranded Total RNA with Ribo-Zero Gold kit.
attributes.tag: ENA first public
attributes.value: 2018-12-01
attributes.tag: ENA last update
attributes.value: 2017-11-02
attributes.tag: External Id
attributes.value: SAMEA104368196
attributes.tag: INSDC center alias
attributes.value: Centre for Discovery Brain Sciences, University of Edinburgh
attributes.tag: INSDC center name
attributes.value: Centre for Discovery Brain Sciences, University of Edinburgh
attributes.tag: INSDC first public
attributes.value: 2018-12-01T17:02:04Z
attributes.tag: INSDC last update
attributes.value: 2017-11-02T15:28:32Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-6167:G1745_HR1
attributes.tag: age
attributes.value: 55
attributes.tag: biopsy site
attributes.value: dermis
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell type
attributes.value: anterior neuronal precursor cell
attributes.tag: clinical history
attributes.value: healthy control
attributes.tag: common name
attributes.value: human
attributes.tag: disease
attributes.value: normal
attributes.tag: individual
attributes.value: G1745
attributes.tag: karyotype
attributes.value: normal
attributes.tag: progenitor cell type
attributes.value: induced pluripotent stem cell
attributes.tag: sample name
attributes.value: E-MTAB-6167:G1745_HR1
attributes.tag: sex
attributes.value: male