characteristics.text: cd34n
characteristics.characteristic: AML status
characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2016-06-22T17:01:37Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2018-03-08T18:23:06Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA2604622
characteristics.characteristic: External Id
characteristics.text: Paresh Vyas group, Molecular haematology Unit, The Weatherall Institute of Molecular Medicine
characteristics.characteristic: INSDC center name
characteristics.text: 2016-06-22T17:01:37Z
characteristics.characteristic: INSDC first public
characteristics.text: 2018-03-08T18:23:06Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS482508
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-2672:Sample #001-34p
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: CD34+
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0005180
characteristics.characteristic: cell type
characteristics.text: human
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_9606
characteristics.characteristic: common name
characteristics.text: Protocols: Mononuclear cells were isolated by Ficoll density gradient. In normal bone marrow samples, CD34+ cells were purified using CD34 Microbead Kit and MACS separation columns (Miltenyi Biotec, Bisley UK). Unseparated, CD34+ and CD34-deplete fractions were frozen in 90% FCS/10% DMSO and stored in liquid nitrogen, and subsequently thawed on the day of the experiment. Antibodies used for lineage-depletion cocktails and in flow cytometry panels in different experiments are shown in Supplemental Experimental Procedures. FACS analysis was carried out on either a Cyan ADP (Dakocytomation, Ely UK) or a BD LSR Fortessa (Becton Dickinson, Oxford UK). Flow sorts were carried out on a BD Aria III SORP (Becton Dickinson, Oxford UK). Total RNA was extracted from 5x103 - 4x104 sorted cells, quantified using the total RNA Pico or Nano Bioanalyzer chip (Agilent, Wokingham UK). All samples had a RIN score of ?7.0. 20pg to 4ng of total RNA was used for cDNA synthesis (SMARTer Ultra Low input RNA kit (Clontech, St Germain-en-Laye France). cDNA libraries were amplified and sequenced on the Illumina HiSeq 2000 and 2500 (paired-end, read length: 50 base pairs; Illumina, Saffron Walden, UK).
characteristics.characteristic: description
characteristics.text: AML
characteristics.characteristic: disease
characteristics.text: #001
characteristics.characteristic: individual
characteristics.text: positive
characteristics.characteristic: leukaemic stem cells
characteristics.text: Homo sapiens
characteristics.characteristic: organism
characteristics.text: Homo sapiens
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_9606
characteristics.characteristic: organism
characteristics.text: CD34+
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0005180
characteristics.characteristic: phenotype
characteristics.text: Homo sapiens
characteristics.characteristic: scientific_name
characteristics.text: Sample #001-34p
characteristics.characteristic: title