title: Sample 481
organism: Homo sapiens
description: Protocols: Purification of iPSC derived dopaminergic neurons was carried out as previously described (Sandor et al., 2017). At sorting each sample was first sorted into each row of a 96 well plate, into 2 ul of smart-seq 2 lysis buffer (0.2% triton x-100 and 2 U/ul RNase inhibitor), so that all samples were on each 96 well plate for single cell RNA-sequencing. After 96 well plate sorting, the rest of the sample was bulk collected for RNA extraction in preparation for bulk RNA-sequencing. RNA from bulk collected FACS sorted dopamine neurons was extracted using an FFPE RNA extraction kit (Qiagen) as per manufacturer's instructions, with minor modifications. RNA integrity analysis was analysed using a 2100 bioanalyzer system and a RNA 6000 pico kit (Agilent). Concentration was obtained and confirmed utilising two methods; the 2100 bioanalyzer system and a Quant-iT RiboGreen RNA kit (Invitrogen), as per manufacturer's instructions. Single cells and RNA extracted from bulks were processed using the Smart-seq2 protocol (Picelli et al., 2013). cDNA samples were prepared for sequencing using the Nextera XT DNA Library Prep Kit (Illumina) with our own in-house indexing primers (Lamble et al., 2013).
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2018-10-17T17:02:31Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2018-10-17T16:25:36Z
attributes.tag: External Id
attributes.value: SAMEA4981245
attributes.tag: INSDC center name
attributes.value: OXFORD
attributes.tag: INSDC first public
attributes.value: 2018-10-17T17:02:31Z
attributes.tag: INSDC last update
attributes.value: 2018-10-17T16:25:36Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-7303:Sample 481
attributes.tag: age
attributes.value: 80
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell type
attributes.value: dopaminergic neuron
attributes.tag: common name
attributes.value: human
attributes.tag: developmental stage
attributes.value: adult
attributes.tag: disease
attributes.value: normal
attributes.tag: genotype
attributes.value: wt/wt
attributes.tag: individual
attributes.value: Control1
attributes.tag: organism part
attributes.value: skin
attributes.tag: progenitor cell type
attributes.value: induced pluripotent stem cell
attributes.tag: sample name
attributes.value: E-MTAB-7303:Sample 481
attributes.tag: scientific_name
attributes.value: Homo sapiens
attributes.tag: sex
attributes.value: male
attributes.tag: single cell well quality
attributes.value: 0
attributes.tag: well information
attributes.value: Plate_based_single_cell