Homo sapiens
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).
AML status
normal
ENA-FIRST-PUBLIC
2016-06-22T17:01:37Z
ENA-LAST-UPDATE
2018-03-08T18:21:42Z
External Id
SAMEA2604604
INSDC center name
Paresh Vyas group, Molecular haematology Unit, The Weatherall Institute of Molecular Medicine
INSDC first public
2016-06-22T17:01:37Z
INSDC last update
2018-03-08T18:21:42Z
INSDC status
public
Submitter Id
E-MTAB-2672:Sample #662-MPP
broker name
ArrayExpress
cell type
MPP
common name
human
disease
normal
individual
#662
leukaemic stem cells
negative
phenotype
MPP
sample name
E-MTAB-2672:Sample #662-MPP
scientific_name
Homo sapiens