characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2023-02-14T08:20:15Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2023-02-14T08:20:15Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA112641131
characteristics.characteristic: External Id
characteristics.text: Division of Stem Cells and Cancer, German Cancer Research Center (DKFZ) and DKFZ-ZMBH Alliance, 69120 Heidelberg, Germany Heidelberg Institute for Stem Cell Technology and Experimental Medicine (HI-STEM gGmbH), 69120 Heidelberg, Germany
characteristics.characteristic: INSDC center name
characteristics.text: 2023-02-14T08:20:15Z
characteristics.characteristic: INSDC first public
characteristics.text: 2023-02-14T08:20:15Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS14633106
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-11976_2:Sample 47
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.characteristic: broker name
characteristics.text: monocyte-like AML
characteristics.characteristic: clinical information
characteristics.text: human
characteristics.characteristic: common name
characteristics.text: Protocols: AML samples were collected from diagnostic peripheral blood (PB) aspirations at the University hospital in Heidelberg in accordance with the Declaration of Helsinki after obtaining written consent from each patient. The project was approved by the Ethics Committee of the Medical Faculty of Heidelberg (S-169/217). PB mononuclear cells (MNCs) were isolated by density gradient centrifugation using Ficoll Paque Plus (GE Healthcare, cat # GE17-1440-03), and stored in liquid nitrogen until further use. Primary AML cells were stained with a total of ten fluorescent cell surface antibodies (see supplementary table X). Cells were sorted into four populations according to CD11b, CD64 and GPR56 expression within the lineage-negative gate. Cells from each population and from lineage-neagtive bulk were sorted directly into RNA extraction buffer (Thermo Fisher, cat # KIT0214), snap-frozen and stored at -80 °C until RNA extraction. RNA extraction and purification of FACS-sorted cells was done using PicoPure RNA Isolation Kit according to manufacturer’s instructions (Thermo Fisher, cat # KIT0214). Whole transcriptome amplification was performed using a modified smart-seq2 protocol (Picelli et al 2014), with 5μl of a modified RT buffer containing 1× SMART First Strand Buffer (Takara Bio Clontech, cat # 639538), 1 mM dithiothreitol (Takara Bio Clontech), 1 μM template switching oligo (IDT), 10 U μl−1 SMARTScribe (Takara Bio Clontech, cat # 639538) and 1 U μl−1 RNasin Plus RNase Inhibitor (Promega, cat # N2615). Tagmentation of cDNA was done using Nextera XT DNA Library Preparation Kit (Illumina, cat # FC-121-1030).
characteristics.characteristic: description
characteristics.text: adult
characteristics.characteristic: developmental stage
characteristics.text: acute myeloid leukemia
characteristics.characteristic: disease
characteristics.text: 41
characteristics.characteristic: individual
characteristics.text: Homo sapiens
characteristics.characteristic: organism
characteristics.text: Homo sapiens
characteristics.characteristic: organism
characteristics.text: blood
characteristics.characteristic: organism part
characteristics.text: non-leukemia stem cell
characteristics.characteristic: phenotype
characteristics.text: Homo sapiens
characteristics.characteristic: scientific_name
characteristics.text: Sample 47
characteristics.characteristic: title