title: Sample #059-mp
organism: Homo sapiens
description: 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).
attributes.tag: AML status
attributes.value: cd34n
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2016-06-22T17:01:37Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2018-03-08T18:21:40Z
attributes.tag: External Id
attributes.value: SAMEA2604598
attributes.tag: INSDC center name
attributes.value: Paresh Vyas group, Molecular haematology Unit, The Weatherall Institute of Molecular Medicine
attributes.tag: INSDC first public
attributes.value: 2016-06-22T17:01:37Z
attributes.tag: INSDC last update
attributes.value: 2018-03-08T18:21:40Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-2672:Sample #059-mp
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell type
attributes.value: CD34-CD244-CD117+
attributes.tag: common name
attributes.value: human
attributes.tag: disease
attributes.value: AML
attributes.tag: individual
attributes.value: #059
attributes.tag: leukaemic stem cells
attributes.value: positive
attributes.tag: phenotype
attributes.value: CD34-CD244-CD117+
attributes.tag: sample name
attributes.value: E-MTAB-2672:Sample #059-mp
attributes.tag: scientific_name
attributes.value: Homo sapiens