E-MTAB-12238:Sample 172

characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC first public

text: public

characteristic: SRA accession

text: E-MTAB-12238:Sample 172

characteristic: Submitter Id

characteristic: WGA

text: ArrayExpress

text: diagnosis

text: acute myeloid leukemia

text: human

text: adult

characteristic: developmental stage

text: acute myeloid leukemia

text: CD34+, CD38-,CD90+/-, CD45Ra-

text: Homo sapiens

text: Homo sapiens

text: bone marrow

characteristic: organism part

text: FACS Aria

characteristic: sample id

text: Homo sapiens

text: male

text: NPM1+ FLT3-ITD+

characteristic: time

text: Sample 172


E-MTAB-12238:Sample 173

status: PUBLIC

tag: attribute

characteristic: INSDC last update

text: public

characteristic: INSDC status

text: E-MTAB-12238:Sample 173

tag: Namespace:Department of Molecular Cell Biology, Weizmann Institute of Science

text: Repli G

text: ArrayExpress

characteristic: broker name

characteristic: cell count

text: diagnosis

characteristic: clinical history

text: acute myeloid leukemia

characteristic: clinical information

text: human

characteristic: common name

characteristic: date

text: Protocols: CD34+ cells were isolated from PB or BM (following mononuclear cell separation) of AML patients by positive selection using anti-human CD34 microbeads. Enriched CD34+ cells were sorted by FACSariaII (BD bioscience, CA, USA) into four subpopulations of HSPCs according to their surface markers and CD33 expression. The sorted fractions were collected either into Iscove's modified Dulbecco's medium (IMDM) +20% fetal bovine serum (FBS) (Biological Industries #02-023-1A) to be further used in the colony forming unit (CFU) assay or into PBS+2%FBS to be used in the xenotransplantation assay. The CFU assay was adjusted to the generation of single cell-derived subclones. HSPCs were seeded in 1% standard methylcellulose medium supplemented with 37% FBS, 1.2% bovine serum albumin (BSA), 1% L-glutamine, 10-6M b-mercaptoethanol, 1 IU/ml of human erythropoietin (EPO), 50 ng/ml of human stem cell factor (SCF), 25 ng/ml of human FLT3 ligand, 10 ng/ml of interleukin 3 (IL-3), 10 ng/ml of interleukin 6 (IL-6), 10 ng/ml of granulocytecolony stimulating factor (G-CSF), 5 ng/ml of granulocyte-macrophage colony-stimulating factor (GM-CSF), 10 ng/ml of human thrombopoietin (TPO), 100 IU ml penicillin and 100 mg/ml streptomycin. Colonies were counted after 14 days and each of them was picked under sterile conditions. Every individual colony was divided into two cell suspensions to perform both genotyping analysis and xenotransplantation into NSG mice. xenograft protocol: NOD.Cg-PrkdcscidIl2rgtm1Wjl/SzJ (NSG) mice (The Jackson Laboratory, Bar Harbor, Maine, USA) were maintained and bred under specific pathogen-free conditions at the Biomedical Core Facility (Faculty of Medicine, the Technion, Haifa, Israel). Twenty-four hours prior to transplantation, 8-12-week-old female NSG mice were sub-lethally irradiated with 225 cGy. Primary human cells were intra-femorally injected. The total number of cells transplanted per mouse depended on the number of viable cells obtained from the BM/PB of a patient [ranged from 1 colony (a single cell-derived subclone) to 106 cells]. The animals were sacrificed 8-30 weeks after transplantation or if tumors developed in the injected knee. The BM was then harvested. Human engraftment was evaluated by flow cytometry using the following anti-human specific antibodies: anti-CD45-APC Cyanine7 (APC-Cy7) (BioLegend #304014 clone H130); anti-CD33-BV510 (BioLegend #366610); anti-CD34-BV421 (BioLegend #343610); anti-CD38-FITC (BD #555459); anti-CD15 APC (BD #551376); anti-CD19 PE-CY7 (BD #557835); Cell viability was obtained using Propidium Iodide - PI (BioLegend #421301). Analyses were performed using FlowJo software v.10 (Treestar, OR, USA), and the expression of markers was assessed and presented as the percentage of stained cells. The threshold for engraftment detection was 0.1% of human CD45+ cells. All flow cytometry analyses were done using an LSR II (BD Bioscience, CA, USA) or FACSariaII (BD Bioscience, CA, USA) device. Engraftment of >5% of hCD45+ cells with the major CD33+ population (>60% of the hCD45+ cells) was classified as overt leukemia; the presence of 0.1-5% of hCD45+ cells or the presence of >5% of hCD45+ cells with <60% of the CD33+ population in the hCD45+ cells was classified as non-overt leukemia; and the presence of <0.1% of human CD45+ (hCD45+) population was considered as non-engraftment. hCD45+ cells were sorted and underwent whole genome amplification (WGA) for further genotyping assays. CD34+ cells were isolated from PB or BM (following mononuclear cell separation) of AML patients by positive selection using anti-human CD34 microbeads. Enriched CD34+ cells were sorted by FACSariaII (BD bioscience, CA, USA) into four subpopulations of HSPCs according to their surface markers and CD33 expression. The sorted fractions were collected either into Iscove's modified Dulbecco's medium (IMDM) +20% fetal bovine serum (FBS) (Biological Industries #02-023-1A) to be further used in the colony forming unit (CFU) assay or into PBS+2%FBS to be used in the xenotransplantation assay. The CFU assay was adjusted to the generation of single cell-derived subclones. HSPCs were seeded in 1% standard methylcellulose medium supplemented with 37% FBS, 1.2% bovine serum albumin (BSA), 1% L-glutamine, 10-6M b-mercaptoethanol, 1 IU/ml of human erythropoietin (EPO), 50 ng/ml of human stem cell factor (SCF), 25 ng/ml of human FLT3 ligand, 10 ng/ml of interleukin 3 (IL-3), 10 ng/ml of interleukin 6 (IL-6), 10 ng/ml of granulocytecolony stimulating factor (G-CSF), 5 ng/ml of granulocyte-macrophage colony-stimulating factor (GM-CSF), 10 ng/ml of human thrombopoietin (TPO), 100 IU ml penicillin and 100 mg/ml streptomycin. Colonies were counted after 14 days and each of them was picked under sterile conditions. Every individual colony was divided into two cell suspensions to perform both genotyping analysis and xenotransplantation into NSG mice One µl of DNA template was added to a hybridization mix together with an MIP pool (final concentration of 0.05pM per probe) in 1x Ampligase buffer (Epicentre). The mix was incubated in a thermal cycler according to the following program: 98°C for 3 minutes, 85°C for 30 minutes, 60°C for 60 minutes and 56°C for 1 or 2 overnight incubations. The product was mixed with (final concentration in brackets): dNTPs (Larova, 15pM), Betaine (375 mM, Sigma-Aldrich,), NAD+ (1 mM, New England Biolabs), additional Ampligase buffer (0.5x), Ampligase (total of 1.25U, Epicentre) and Phusion HF (0.16U, New England Biolabs). The mix was incubated at 56°C for 60 minutes followed by 72°C for 20 minutes. Enzymatic digestion of linear probes was performed by adding Exonuclease I (4U, New England Biolabs) and Exonuclease III (25U, New England Biolabs). The mix was incubated at 37°C for 2 hours, followed by 80°C for 20 minutes. The final product was amplified using iProof HF Master Mix (Biorad). Samples were pooled and concentrated using AMPure XP beads (Beckman Coulter) at 1.3x volumetric concentration, size-selected (190-370bp) using Blue Pippin (Sage scientific), and sequenced in either NextSeq or Novaseq6000 (Illumina) 2X151bp paired-end run using custom primers as described in the past(Biezuner et al., 2022)

characteristic: description

text: adult

text: acute myeloid leukemia

characteristic: disease

characteristic: individual

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: blood

text: FACS Aria

characteristic: protocol

text: Homo sapiens

text: male

characteristic: sex

text: NPM1+ FLT3-ITD+; CD45 positive

characteristic: subclone

text: Sample 173

characteristic: title

text: human CD45 positive out of mouse bone marrow 6

characteristic: xenograft


E-MTAB-12238:Sample 174

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text: Department of Molecular Cell Biology, Weizmann Institute of Science

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text: E-MTAB-12238:Sample 174

characteristic: Submitter Id

text: Repli G

characteristic: WGA

text: ArrayExpress

text: diagnosis

text: acute myeloid leukemia

text: human

text: adult

characteristic: developmental stage

text: acute myeloid leukemia

text: Homo sapiens

text: Homo sapiens

text: blood

characteristic: organism part

text: FACS Aria

characteristic: sample id

text: Homo sapiens

text: male

text: NPM1+ FLT3-ITD+; CD45 positive

characteristic: time

text: Sample 174


E-MTAB-12238:Sample 175

text: public

characteristic: INSDC status

text: E-MTAB-12238:Sample 175

text: Repli G

text: ArrayExpress

characteristic: broker name

text: diagnosis

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text: acute myeloid leukemia

characteristic: clinical information

text: human

characteristic: common name

characteristic: date

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text: adult

text: acute myeloid leukemia

characteristic: disease

characteristic: individual

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: blood

text: FACS Aria

characteristic: protocol

text: Homo sapiens

text: male

characteristic: sex

text: NPM1+ FLT3-ITD+; CD45 positive

characteristic: subclone

text: Sample 175

text: human CD45 positive out of mouse bone marrow 102


E-MTAB-12238:Sample 176

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text: Repli G

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text: ArrayExpress

text: diagnosis

text: acute myeloid leukemia

text: human

text: adult

characteristic: developmental stage

text: acute myeloid leukemia

text: Homo sapiens

text: Homo sapiens

text: blood

text: FACS Aria

text: Homo sapiens

text: male

text: NPM1+ FLT3-ITD+; CD45 positive

text: Sample 176

characteristic: title

characteristic: xenograft


E-MTAB-12238:Sample 177

text: public

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text: ArrayExpress

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text: common lymphoid progenitor

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text: acute myeloid leukemia

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text: adult

text: acute myeloid leukemia

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text: CD34+, CD38-,CD90+/-, CD45Ra+

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text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: bone marrow

characteristic: organism part

text: FACS Aria

characteristic: sample id

text: Homo sapiens

text: male

text: NPM1+ FLT3-ITD+

characteristic: time

text: Sample 177


E-MTAB-12238:Sample 178

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text: ArrayExpress

text: common lymphoid progenitor

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text: acute myeloid leukemia

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text: acute myeloid leukemia

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text: FACS Aria

characteristic: protocol

text: Homo sapiens

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characteristic: sex

text: NPM1+ FLT3-ITD+

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text: Sample 178


E-MTAB-12238:Sample 179

text: public

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text: Repli G

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characteristic: broker name

text: common myeloid progenitor

text: relapse

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characteristic: clinical information

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text: adult

text: acute myeloid leukemia

characteristic: disease

text: CD34+, CD38+, CD10-, CD7-, CD135(FLT3)+, CD45Ra-

characteristic: immunophenotype

characteristic: individual

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: bone marrow

characteristic: organism part

text: FACS Aria

text: Homo sapiens

text: male

text: BCR-ABL/NPM1-/FLT3 ITD-/DNMT3A-

text: Sample 179

characteristic: title


E-MTAB-12238:Sample 18

characteristic: ENA-CHECKLIST

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characteristic: Submitter Id

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characteristic: time

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text: 1:10 human CD45 positive out of mouse bone marrow 80

characteristic: xenograft


E-MTAB-12238:Sample 180

text: public

characteristic: INSDC status

text: E-MTAB-12238:Sample 180

text: Repli G

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text: human

characteristic: common name

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text: adult

text: acute myeloid leukemia

characteristic: disease

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characteristic: individual

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: bone marrow

text: FACS Aria

characteristic: protocol

text: Homo sapiens

text: male

characteristic: sex

text: BCR-ABL/NPM1-/FLT3 ITD-/DNMT3A-

characteristic: subclone

text: Sample 180


E-MTAB-12238:Sample 181

characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

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characteristic: INSDC center name

characteristic: INSDC first public

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text: public

characteristic: SRA accession

text: E-MTAB-12238:Sample 181

characteristic: Submitter Id

characteristic: WGA

text: ArrayExpress

text: hematopoietic multipotent progenitor cell

text: diagnosis

text: acute myeloid leukemia

text: human

text: adult

characteristic: developmental stage

text: acute myeloid leukemia

text: CD34+, CD38-,CD90+/-, CD45Ra-

text: Homo sapiens

text: Homo sapiens

text: bone marrow

characteristic: organism part

text: FACS Aria

characteristic: sample id

text: Homo sapiens

text: male

text: NPM1+ FLT3-ITD+

text: Sample 181

characteristic: title


E-MTAB-12238:Sample 182

text: public

characteristic: INSDC status

text: E-MTAB-12238:Sample 182

text: ArrayExpress

characteristic: broker name

text: diagnosis

characteristic: clinical history

text: acute myeloid leukemia

characteristic: clinical information

text: human

characteristic: common name

characteristic: date

characteristic: description

text: adult

text: acute myeloid leukemia

characteristic: disease

text: CD34+, CD38-,CD90+/-, CD45Ra-

characteristic: immunophenotype

characteristic: individual

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: bone marrow

text: FACS Aria

characteristic: protocol

text: Homo sapiens

text: male

text: NPM1+ FLT3-ITD+

characteristic: time

text: Sample 182


E-MTAB-12238:Sample 183

characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC first public

characteristic: INSDC last update

text: public

characteristic: SRA accession

text: E-MTAB-12238:Sample 183

characteristic: Submitter Id

text: Repli G

characteristic: WGA

text: ArrayExpress

characteristic: cell count

text: common lymphoid progenitor

text: diagnosis

text: acute myeloid leukemia

text: human

text: adult

characteristic: developmental stage

text: acute myeloid leukemia

text: CD34+, CD38-,CD90+/-, CD45Ra+

text: Homo sapiens

text: Homo sapiens

text: blood

text: FACS Aria

text: Homo sapiens

text: male

characteristic: sex

text: NPM1+ FLT3-ITD+

characteristic: subclone

text: Sample 183

text: 1:10 human CD45 positive out of mouse bone marrow 6

characteristic: xenograft


E-MTAB-12238:Sample 184

text: public

characteristic: INSDC status

text: E-MTAB-12238:Sample 184

text: Repli G

text: ArrayExpress

characteristic: broker name

text: common lymphoid progenitor

text: diagnosis

characteristic: clinical history

text: acute myeloid leukemia

characteristic: clinical information

text: human

characteristic: common name

characteristic: date

characteristic: description

text: adult

text: acute myeloid leukemia

characteristic: disease

text: CD34+, CD38-,CD90+/-, CD45Ra+

characteristic: immunophenotype

characteristic: individual

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: blood

characteristic: organism part

text: FACS Aria

characteristic: protocol

characteristic: sample id

text: Homo sapiens

text: male

text: NPM1+ FLT3-ITD+

text: Sample 184

characteristic: title


E-MTAB-12238:Sample 185

characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

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characteristic: INSDC center name

characteristic: INSDC first public

characteristic: INSDC last update

text: public

characteristic: SRA accession

text: E-MTAB-12238:Sample 185

characteristic: Submitter Id

text: Repli G

characteristic: WGA

text: ArrayExpress

text: common lymphoid progenitor

text: diagnosis

text: acute myeloid leukemia

text: human

text: adult

characteristic: developmental stage

text: acute myeloid leukemia

text: CD34+, CD38-,CD90+/-, CD45Ra+

text: Homo sapiens

text: Homo sapiens

text: blood

text: FACS Aria

text: Homo sapiens

text: male

characteristic: sex

text: NPM1+ FLT3-ITD+

characteristic: subclone

characteristic: time

text: Sample 185

text: 1:10 human CD45 positive out of mouse bone marrow 102


E-MTAB-12238:Sample 186

text: public

characteristic: INSDC status

text: E-MTAB-12238:Sample 186

text: Repli G

text: ArrayExpress

characteristic: broker name

text: common lymphoid progenitor

text: diagnosis

characteristic: clinical history

text: acute myeloid leukemia

characteristic: clinical information

text: human

characteristic: common name

characteristic: date

characteristic: description

text: adult

text: acute myeloid leukemia

characteristic: disease

text: CD34+, CD38-,CD90+/-, CD45Ra+

characteristic: immunophenotype

characteristic: individual

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: blood

characteristic: organism part

text: FACS Aria

characteristic: protocol

text: Homo sapiens

text: male

text: NPM1+ FLT3-ITD+

text: Sample 186

characteristic: xenograft


E-MTAB-12238:Sample 187

characteristic: ENA-CHECKLIST

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characteristic: INSDC center name

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text: public

characteristic: SRA accession

text: E-MTAB-12238:Sample 187

characteristic: Submitter Id

characteristic: WGA

text: ArrayExpress

text: common lymphoid progenitor

text: diagnosis

text: acute myeloid leukemia

text: human

text: adult

characteristic: developmental stage

text: acute myeloid leukemia

text: CD34+, CD38-,CD90+/-, CD45Ra+

text: Homo sapiens

text: Homo sapiens

text: bone marrow

text: FACS Aria

characteristic: sample id

text: Homo sapiens

text: male

characteristic: sex

text: NPM1+ FLT3-ITD+

text: Sample 187

characteristic: title


E-MTAB-12238:Sample 188

text: public

characteristic: INSDC status

text: E-MTAB-12238:Sample 188

text: ArrayExpress

characteristic: broker name

text: common lymphoid progenitor

text: diagnosis

characteristic: clinical history

text: acute myeloid leukemia

characteristic: clinical information

text: human

characteristic: common name

characteristic: date

characteristic: description

text: adult

text: acute myeloid leukemia

characteristic: disease

text: CD34+, CD38-,CD90+/-, CD45Ra+

characteristic: immunophenotype

characteristic: individual

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: bone marrow

characteristic: organism part

text: FACS Aria

text: Homo sapiens

text: male

text: NPM1+ FLT3-ITD+

characteristic: subclone

characteristic: time

text: Sample 188


E-MTAB-12238:Sample 189

characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC first public

characteristic: INSDC last update

text: public

characteristic: SRA accession

text: E-MTAB-12238:Sample 189

characteristic: Submitter Id

characteristic: WGA

text: ArrayExpress

text: diagnosis

text: acute myeloid leukemia

text: human

text: adult

characteristic: developmental stage

text: acute myeloid leukemia

text: CD34+, CD38-,CD90+/-, CD45Ra-

text: Homo sapiens

text: Homo sapiens

text: bone marrow

text: FACS Aria

characteristic: protocol

characteristic: sample id

text: Homo sapiens

text: male

text: NPM1+ FLT3-ITD+

text: Sample 189


E-MTAB-12238:Sample 19

text: public

characteristic: INSDC status

text: E-MTAB-12238:Sample 19

text: Repli G

text: ArrayExpress

characteristic: broker name

text: common myeloid progenitor

text: diagnosis

characteristic: clinical history

text: acute myeloid leukemia B phenotype

characteristic: clinical information

text: human

characteristic: common name

characteristic: date

characteristic: description

text: adult

text: acute myeloid leukemia

characteristic: disease

characteristic: immunophenotype

characteristic: individual

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: bone marrow

text: FACS Aria - H4100 colony

text: Homo sapiens

text: male

characteristic: sex

text: BCR-ABL/NPM1-/FLT3 ITD-/DNMT3A-

text: Sample 19

characteristic: title


E-MTAB-12238:Sample 190

characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC first public

characteristic: INSDC last update

text: public

characteristic: SRA accession

text: E-MTAB-12238:Sample 190

characteristic: Submitter Id

characteristic: WGA

text: ArrayExpress

text: diagnosis

text: acute myeloid leukemia

text: human

text: adult

text: acute myeloid leukemia

text: CD34+, CD38-,CD90+/-, CD45Ra-

text: Homo sapiens

text: Homo sapiens

text: bone marrow

characteristic: organism part

text: FACS Aria

text: Homo sapiens

text: male

text: NPM1+ FLT3-ITD+

characteristic: subclone

characteristic: time

text: Sample 190


E-MTAB-12238:Sample 191

text: public

characteristic: INSDC status

text: E-MTAB-12238:Sample 191

text: Repli G

text: ArrayExpress

characteristic: broker name

text: diagnosis

characteristic: clinical history

text: acute myeloid leukemia

characteristic: clinical information

text: human

characteristic: common name

characteristic: date

characteristic: description

text: adult

characteristic: developmental stage

text: acute myeloid leukemia

characteristic: disease

characteristic: individual

text: Homo sapiens

text: Homo sapiens

text: blood

text: FACS Aria

characteristic: protocol

characteristic: sample id

text: Homo sapiens

text: male

text: NPM1+ FLT3-ITD+; CD45 positive

text: Sample 191

text: human CD45 positive out of mouse bone marrow 77

characteristic: xenograft


E-MTAB-12238:Sample 192

characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC first public

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text: public

characteristic: SRA accession

text: E-MTAB-12238:Sample 192

characteristic: Submitter Id

text: Repli G

characteristic: WGA

text: ArrayExpress

text: diagnosis

text: acute myeloid leukemia

text: human

text: adult

text: acute myeloid leukemia

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: blood

text: FACS Aria

text: Homo sapiens

text: male

characteristic: sex

text: NPM1+ FLT3-ITD+; CD45 positive

text: Sample 192

characteristic: title


E-MTAB-12238:Sample 193

text: public

characteristic: INSDC status

text: E-MTAB-12238:Sample 193

text: Repli G

text: ArrayExpress

characteristic: broker name

text: diagnosis

characteristic: clinical history

text: acute myeloid leukemia

characteristic: clinical information

text: human

characteristic: common name

characteristic: date

characteristic: description

text: adult

characteristic: developmental stage

text: acute myeloid leukemia

characteristic: disease

characteristic: individual

text: Homo sapiens

text: Homo sapiens

text: blood

characteristic: organism part

text: FACS Aria

text: Homo sapiens

text: male

text: NPM1+ FLT3-ITD+; CD45 positive

characteristic: subclone

characteristic: time

text: Sample 193

text: human CD45 positive out of mouse bone marrow 103


E-MTAB-12238:Sample 194

characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: External Id

characteristic: INSDC center name

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characteristic: INSDC last update

text: public

characteristic: SRA accession

text: E-MTAB-12238:Sample 194

characteristic: Submitter Id

text: Repli G

characteristic: WGA

text: ArrayExpress

text: diagnosis

text: acute myeloid leukemia

text: human

text: adult

text: acute myeloid leukemia

text: Homo sapiens

text: Homo sapiens

text: blood

text: FACS Aria

characteristic: protocol

characteristic: sample id

text: Homo sapiens

text: male

text: NPM1+ FLT3-ITD+; CD45 positive

text: Sample 194

characteristic: xenograft


E-MTAB-12238:Sample 195

text: public

characteristic: INSDC status

text: E-MTAB-12238:Sample 195

text: ArrayExpress

characteristic: broker name

text: common lymphoid progenitor

text: diagnosis

characteristic: clinical history

text: acute myeloid leukemia

characteristic: clinical information

text: human

characteristic: common name

characteristic: date

characteristic: description

text: adult

characteristic: developmental stage

text: acute myeloid leukemia

characteristic: disease

text: CD34+, CD38-,CD90+/-, CD45Ra+

characteristic: immunophenotype

characteristic: individual

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: bone marrow

text: FACS Aria

text: Homo sapiens

text: male

characteristic: sex

text: NPM1+ FLT3-ITD+

text: Sample 195

characteristic: title


E-MTAB-12238:Sample 196

characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC first public

characteristic: INSDC last update

text: public

characteristic: SRA accession

text: E-MTAB-12238:Sample 196

characteristic: Submitter Id

characteristic: WGA

text: ArrayExpress

text: common lymphoid progenitor

text: diagnosis

text: acute myeloid leukemia

text: human

text: adult

text: acute myeloid leukemia

text: CD34+, CD38-,CD90+/-, CD45Ra+

text: Homo sapiens

text: Homo sapiens

text: bone marrow

characteristic: organism part

text: FACS Aria

characteristic: protocol

text: Homo sapiens

text: male

text: NPM1+ FLT3-ITD+

characteristic: subclone

characteristic: time

text: Sample 196


E-MTAB-12238:Sample 197

text: public

characteristic: INSDC status

text: E-MTAB-12238:Sample 197

text: ArrayExpress

characteristic: broker name

text: common lymphoid progenitor

text: diagnosis

characteristic: clinical history

text: acute myeloid leukemia

characteristic: clinical information

text: human

characteristic: common name

characteristic: date

characteristic: description

text: adult

characteristic: developmental stage

text: acute myeloid leukemia

characteristic: disease

text: CD34+, CD38-,CD90+/-, CD45Ra+

characteristic: immunophenotype

characteristic: individual

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: bone marrow

text: FACS Aria

characteristic: sample id

text: Homo sapiens

text: male

text: NPM1+ FLT3-ITD+

text: Sample 197


E-MTAB-12238:Sample 198

characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC first public

characteristic: INSDC last update

text: public

characteristic: SRA accession

text: E-MTAB-12238:Sample 198

characteristic: Submitter Id

characteristic: WGA

text: ArrayExpress

text: common lymphoid progenitor

text: diagnosis

text: acute myeloid leukemia

text: human

text: adult

text: acute myeloid leukemia

text: CD34+, CD38-,CD90+/-, CD45Ra+

text: Homo sapiens

text: Homo sapiens

text: bone marrow

characteristic: organism part

text: FACS Aria

text: Homo sapiens

text: male

characteristic: sex

text: NPM1+ FLT3-ITD+

text: Sample 198

characteristic: title


E-MTAB-12238:Sample 199

text: public

characteristic: INSDC status

text: E-MTAB-12238:Sample 199

text: Repli G

text: ArrayExpress

characteristic: broker name

text: diagnosis

characteristic: clinical history

text: acute myeloid leukemia

characteristic: clinical information

text: human

characteristic: common name

characteristic: date

characteristic: description

text: adult

characteristic: developmental stage

text: acute myeloid leukemia

characteristic: disease

text: CD34+, CD38-,CD90+/-, CD45Ra-

characteristic: immunophenotype

text: Homo sapiens

text: Homo sapiens

text: blood

text: FACS Aria

characteristic: protocol

text: Homo sapiens

text: male

text: NPM1+ FLT3-ITD+

characteristic: subclone

characteristic: time

text: Sample 199

text: 1:10 human CD45 positive out of mouse bone marrow 77

characteristic: xenograft


E-MTAB-12238:Sample 2

characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC first public

characteristic: INSDC last update

text: public

characteristic: SRA accession

text: E-MTAB-12238:Sample 2

characteristic: Submitter Id

text: ArrayExpress

text: diagnosis

text: acute myeloid leukemia

text: human

text: adult

text: acute myeloid leukemia

characteristic: individual

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: bone marrow

characteristic: organism part

text: DNA Purification from BM

characteristic: sample id

text: Homo sapiens

text: male

text: NPM1+ FLT3-ITD+

text: Sample 2


E-MTAB-12238:Sample 20

text: public

characteristic: INSDC status

text: E-MTAB-12238:Sample 20

text: Repli G

characteristic: WGA

text: ArrayExpress

characteristic: broker name

text: common myeloid progenitor

text: diagnosis

characteristic: clinical history

text: acute myeloid leukemia B phenotype

characteristic: clinical information

text: human

characteristic: common name

characteristic: date

characteristic: description

text: adult

characteristic: developmental stage

text: acute myeloid leukemia

characteristic: disease

characteristic: immunophenotype

text: Homo sapiens

text: Homo sapiens

text: bone marrow

text: FACS Aria - H4100 colony

characteristic: protocol

text: Homo sapiens

text: male

characteristic: sex

text: BCR-ABL/NPM1-/FLT3 ITD-/DNMT3A-

text: Sample 20

characteristic: title


E-MTAB-12238:Sample 200

characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC first public

characteristic: INSDC last update

text: public

characteristic: SRA accession

text: E-MTAB-12238:Sample 200

characteristic: Submitter Id

text: Repli G

text: ArrayExpress

text: diagnosis

text: acute myeloid leukemia

text: human

text: adult

text: acute myeloid leukemia

text: CD34+, CD38-,CD90+/-, CD45Ra-

characteristic: individual

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: blood

text: FACS Aria

text: Homo sapiens

text: male

text: NPM1+ FLT3-ITD+

characteristic: subclone

characteristic: time

text: Sample 200

characteristic: xenograft


E-MTAB-12238:Sample 201

text: public

characteristic: INSDC status

text: E-MTAB-12238:Sample 201

text: Repli G

characteristic: WGA

text: ArrayExpress

characteristic: broker name

text: granulocyte monocyte progenitor cell

text: diagnosis

characteristic: clinical history

text: acute myeloid leukemia B phenotype

characteristic: clinical information

text: human

characteristic: common name

characteristic: date

characteristic: description

text: adult

characteristic: developmental stage

text: acute myeloid leukemia

characteristic: disease

text: CD34+, CD38+, CD10-, CD7-, CD135(FLT3)+, CD45Ra+

characteristic: immunophenotype

text: Homo sapiens

text: Homo sapiens

text: bone marrow

characteristic: organism part

text: FACS Aria

characteristic: protocol

characteristic: sample id

text: Homo sapiens

text: male

text: BCR-ABL/NPM1-/FLT3 ITD-/DNMT3A-

text: Sample 201


E-MTAB-12238:Sample 202

characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC first public

characteristic: INSDC last update

text: public

characteristic: SRA accession

text: E-MTAB-12238:Sample 202

characteristic: Submitter Id

text: Repli G

text: ArrayExpress

text: granulocyte monocyte progenitor cell

text: diagnosis

text: acute myeloid leukemia B phenotype

text: human

text: adult

text: acute myeloid leukemia

characteristic: individual

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: bone marrow

text: FACS Aria

text: Homo sapiens

text: male

characteristic: sex

text: BCR-ABL/NPM1-/FLT3 ITD-/DNMT3A-

characteristic: subclone

text: Sample 202

characteristic: title


E-MTAB-12238:Sample 203

text: public

characteristic: INSDC status

text: E-MTAB-12238:Sample 203

text: Repli G

characteristic: WGA

text: ArrayExpress

characteristic: broker name

text: common lymphoid progenitor

text: diagnosis

characteristic: clinical history

text: acute myeloid leukemia

characteristic: clinical information

text: human

characteristic: common name

characteristic: date

characteristic: description

text: adult

characteristic: developmental stage

text: acute myeloid leukemia

characteristic: disease

text: CD34+, CD38-,CD90+/-, CD45Ra+

characteristic: immunophenotype

text: Homo sapiens

text: Homo sapiens

text: blood

characteristic: organism part

text: FACS Aria

text: Homo sapiens

text: male

text: NPM1+ FLT3-ITD+

characteristic: time

text: Sample 203

text: 1:10 human CD45 positive out of mouse bone marrow 103

characteristic: xenograft


E-MTAB-12238:Sample 204

characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC first public

characteristic: INSDC last update

text: public

characteristic: SRA accession

text: E-MTAB-12238:Sample 204

characteristic: Submitter Id

text: Repli G

text: ArrayExpress

text: common lymphoid progenitor

text: diagnosis

text: acute myeloid leukemia

text: human

text: adult

text: acute myeloid leukemia

text: CD34+, CD38-,CD90+/-, CD45Ra+

characteristic: individual

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: blood

text: FACS Aria

characteristic: protocol

characteristic: sample id

text: Homo sapiens

text: male

characteristic: sex

text: NPM1+ FLT3-ITD+

text: Sample 204


E-MTAB-12238:Sample 205

text: public

characteristic: INSDC status

text: E-MTAB-12238:Sample 205

characteristic: WGA

text: ArrayExpress

characteristic: broker name

text: common lymphoid progenitor

text: diagnosis

characteristic: clinical history

text: acute myeloid leukemia

characteristic: clinical information

text: human

characteristic: common name

characteristic: date

characteristic: description

text: adult

characteristic: developmental stage

text: acute myeloid leukemia

characteristic: disease

text: CD34+, CD38-,CD90+/-, CD45Ra+

characteristic: immunophenotype

text: Homo sapiens

text: Homo sapiens

text: bone marrow

characteristic: organism part

text: FACS Aria

text: Homo sapiens

text: male

text: NPM1+ FLT3-ITD+

characteristic: subclone

text: Sample 205

characteristic: title


E-MTAB-12238:Sample 206

characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC first public

characteristic: INSDC last update

text: public

characteristic: SRA accession

text: E-MTAB-12238:Sample 206

characteristic: Submitter Id

text: ArrayExpress

text: common lymphoid progenitor

text: diagnosis

text: acute myeloid leukemia

text: human

text: adult

text: acute myeloid leukemia

text: CD34+, CD38-,CD90+/-, CD45Ra+

characteristic: individual

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: bone marrow

text: FACS Aria

characteristic: protocol

text: Homo sapiens

text: male

text: NPM1+ FLT3-ITD+

characteristic: time

text: Sample 206


E-MTAB-12238:Sample 207

text: public

characteristic: INSDC status

text: E-MTAB-12238:Sample 207

characteristic: WGA

text: ArrayExpress

characteristic: broker name

text: common lymphoid progenitor

text: diagnosis

characteristic: clinical history

text: acute myeloid leukemia

characteristic: clinical information

text: human

characteristic: common name

characteristic: date

characteristic: description

text: adult

characteristic: developmental stage

text: acute myeloid leukemia

characteristic: disease

text: CD34+, CD38-,CD90+/-, CD45Ra+

characteristic: immunophenotype

text: Homo sapiens

text: Homo sapiens

text: bone marrow

text: FACS Aria

characteristic: sample id

text: Homo sapiens

text: male

characteristic: sex

text: NPM1+ FLT3-ITD+

text: Sample 207


E-MTAB-12238:Sample 208

characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC first public

characteristic: INSDC last update

text: public

characteristic: SRA accession

text: E-MTAB-12238:Sample 208

characteristic: Submitter Id

text: ArrayExpress

text: common lymphoid progenitor

text: diagnosis

text: acute myeloid leukemia

text: human

text: adult

text: acute myeloid leukemia

text: CD34+, CD38-,CD90+/-, CD45Ra+

characteristic: individual

text: Homo sapiens

text: Homo sapiens

text: bone marrow

characteristic: organism part

text: FACS Aria

text: Homo sapiens

text: male

text: NPM1+ FLT3-ITD+

characteristic: subclone

text: Sample 208

characteristic: title


E-MTAB-12238:Sample 209

text: public

characteristic: INSDC status

text: E-MTAB-12238:Sample 209

text: Repli G

characteristic: WGA

text: ArrayExpress

characteristic: broker name

text: diagnosis

characteristic: clinical history

text: acute myeloid leukemia

characteristic: clinical information

text: human

characteristic: common name

characteristic: date

characteristic: description

text: adult

characteristic: developmental stage

text: acute myeloid leukemia

characteristic: disease

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: blood

text: FACS Aria

characteristic: protocol

text: Homo sapiens

text: male

text: NPM1+ FLT3-ITD+; CD45 positive

characteristic: time

text: Sample 209

text: human CD45 positive out of mouse bone marrow 78

characteristic: xenograft


E-MTAB-12238:Sample 21

characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC first public

characteristic: INSDC last update

text: public

characteristic: SRA accession

text: E-MTAB-12238:Sample 21

characteristic: Submitter Id

text: Repli G

text: ArrayExpress

text: diagnosis

text: acute myeloid leukemia B phenotype

text: human

text: adult

text: acute myeloid leukemia

text: CD34+, CD38-,CD90+/-, CD45Ra-

characteristic: immunophenotype

characteristic: individual

text: Homo sapiens

text: Homo sapiens

text: bone marrow

characteristic: organism part

text: FACS Aria - H4100 colony

characteristic: sample id

text: Homo sapiens

text: male

characteristic: sex

text: BCR-ABL/NPM1-/FLT3 ITD-/DNMT3A-

text: Sample 21


E-MTAB-12238:Sample 210

text: public

characteristic: INSDC status

text: E-MTAB-12238:Sample 210

text: Repli G

characteristic: WGA

text: ArrayExpress

characteristic: broker name

text: diagnosis

characteristic: clinical history

text: acute myeloid leukemia

characteristic: clinical information

text: human

characteristic: common name

characteristic: date

characteristic: description

text: adult

characteristic: developmental stage

text: acute myeloid leukemia

characteristic: disease

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: blood

text: FACS Aria

characteristic: protocol

text: Homo sapiens

text: male

text: NPM1+ FLT3-ITD+; CD45 positive

characteristic: subclone

text: Sample 210

characteristic: title


E-MTAB-12238:Sample 211

characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC first public

characteristic: INSDC last update

text: public

characteristic: SRA accession

text: E-MTAB-12238:Sample 211

characteristic: Submitter Id

text: Repli G

text: ArrayExpress

text: diagnosis

text: acute myeloid leukemia

text: human

text: adult

text: acute myeloid leukemia

characteristic: individual

text: Homo sapiens

text: Homo sapiens

text: blood

characteristic: organism part

text: FACS Aria

text: Homo sapiens

text: male

text: NPM1+ FLT3-ITD+; CD45 positive

characteristic: time

text: Sample 211

text: human CD45 positive out of mouse bone marrow 104

characteristic: xenograft


E-MTAB-12238:Sample 212

text: public

characteristic: INSDC status

text: E-MTAB-12238:Sample 212

text: Repli G

characteristic: WGA

text: ArrayExpress

characteristic: broker name

text: diagnosis

characteristic: clinical history

text: acute myeloid leukemia

characteristic: clinical information

text: human

characteristic: common name

characteristic: date

characteristic: description

text: adult

characteristic: developmental stage

text: acute myeloid leukemia

characteristic: disease

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: blood

text: FACS Aria

characteristic: sample id

text: Homo sapiens

text: male

characteristic: sex

text: NPM1+ FLT3-ITD+; CD45 positive

text: Sample 212


E-MTAB-12238:Sample 213

characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC first public

characteristic: INSDC last update

text: public

characteristic: SRA accession

text: E-MTAB-12238:Sample 213

characteristic: Submitter Id

text: ArrayExpress

text: common lymphoid progenitor

text: diagnosis

text: acute myeloid leukemia

text: human

text: adult

text: acute myeloid leukemia

text: CD34+, CD38-,CD90+/-, CD45Ra+

characteristic: immunophenotype

characteristic: individual

text: Homo sapiens

text: Homo sapiens

text: bone marrow

text: FACS Aria

characteristic: protocol

text: Homo sapiens

text: male

text: NPM1+ FLT3-ITD+

characteristic: subclone

text: Sample 213

characteristic: title


E-MTAB-12238:Sample 214

text: public

characteristic: INSDC status

text: E-MTAB-12238:Sample 214

characteristic: WGA

text: ArrayExpress

characteristic: broker name

text: common lymphoid progenitor

text: diagnosis

characteristic: clinical history

text: acute myeloid leukemia

characteristic: clinical information

text: human

characteristic: common name

characteristic: date

characteristic: description

text: adult

characteristic: developmental stage

text: acute myeloid leukemia

characteristic: disease

text: CD34+, CD38-,CD90+/-, CD45Ra+

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: bone marrow

characteristic: organism part

text: FACS Aria

text: Homo sapiens

text: male

text: NPM1+ FLT3-ITD+

characteristic: time

text: Sample 214


E-MTAB-12238:Sample 215

characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC first public

characteristic: INSDC last update

text: public

characteristic: SRA accession

text: E-MTAB-12238:Sample 215

characteristic: Submitter Id

text: ArrayExpress

text: diagnosis

text: acute myeloid leukemia

text: human

text: adult

text: acute myeloid leukemia

characteristic: individual

text: Homo sapiens

text: Homo sapiens

text: bone marrow

text: DNA Purification from BM

characteristic: sample id

text: Homo sapiens

text: female

characteristic: sex

text: Sample 215


E-MTAB-12238:Sample 216

text: public

characteristic: INSDC status

text: E-MTAB-12238:Sample 216

text: ArrayExpress

characteristic: broker name

text: diagnosis

characteristic: clinical history

text: acute myeloid leukemia

characteristic: clinical information

text: human

characteristic: common name

characteristic: date

characteristic: description

text: adult

characteristic: developmental stage

text: acute myeloid leukemia

text: Homo sapiens

text: Homo sapiens

text: bone marrow

text: DNA Purification from BM

characteristic: protocol

text: Homo sapiens

text: female

characteristic: subclone

text: Sample 216

characteristic: title


E-MTAB-12238:Sample 217

characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC first public

characteristic: INSDC last update

text: public

text: E-MTAB-12238:Sample 217

text: Repli G

characteristic: WGA

text: ArrayExpress

text: common myeloid progenitor

text: diagnosis

text: acute myeloid leukemia

text: human

text: adult

text: acute myeloid leukemia

characteristic: disease

characteristic: immunophenotype

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: blood

characteristic: organism part

text: FACS Aria

text: Homo sapiens

text: male

text: NPM1+ FLT3-ITD+

characteristic: time

text: Sample 217

text: 1:10 human CD45 positive out of mouse bone marrow 78

characteristic: xenograft


E-MTAB-12238:Sample 218

text: public

characteristic: INSDC status

characteristic: SRA accession

text: E-MTAB-12238:Sample 218

characteristic: Submitter Id

text: Repli G

text: ArrayExpress

characteristic: broker name

text: common myeloid progenitor

text: diagnosis

characteristic: clinical history

text: acute myeloid leukemia

characteristic: clinical information

text: human

characteristic: common name

characteristic: date

text: Protocols: CD34+ cells were isolated from PB or BM (following mononuclear cell separation) of AML patients by positive selection using anti-human CD34 microbeads. Enriched CD34+ cells were sorted by FACSariaII (BD bioscience, CA, USA) into four subpopulations of HSPCs according to their surface markers and CD33 expression. The sorted fractions were collected either into Iscove's modified Dulbecco's medium (IMDM) +20% fetal bovine serum (FBS) (Biological Industries #02-023-1A) to be further used in the colony forming unit (CFU) assay or into PBS+2%FBS to be used in the xenotransplantation assay. The CFU assay was adjusted to the generation of single cell-derived subclones. HSPCs were seeded in 1% standard methylcellulose medium supplemented with 37% FBS, 1.2% bovine serum albumin (BSA), 1% L-glutamine, 10-6M b-mercaptoethanol, 1 IU/ml of human erythropoietin (EPO), 50 ng/ml of human stem cell factor (SCF), 25 ng/ml of human FLT3 ligand, 10 ng/ml of interleukin 3 (IL-3), 10 ng/ml of interleukin 6 (IL-6), 10 ng/ml of granulocytecolony stimulating factor (G-CSF), 5 ng/ml of granulocyte-macrophage colony-stimulating factor (GM-CSF), 10 ng/ml of human thrombopoietin (TPO), 100 IU ml penicillin and 100 mg/ml streptomycin. Colonies were counted after 14 days and each of them was picked under sterile conditions. Every individual colony was divided into two cell suspensions to perform both genotyping analysis and xenotransplantation into NSG mice. xenograft protocol: NOD.Cg-PrkdcscidIl2rgtm1Wjl/SzJ (NSG) mice (The Jackson Laboratory, Bar Harbor, Maine, USA) were maintained and bred under specific pathogen-free conditions at the Biomedical Core Facility (Faculty of Medicine, the Technion, Haifa, Israel). Twenty-four hours prior to transplantation, 8-12-week-old female NSG mice were sub-lethally irradiated with 225 cGy. Primary human cells were intra-femorally injected. The total number of cells transplanted per mouse depended on the number of viable cells obtained from the BM/PB of a patient [ranged from 1 colony (a single cell-derived subclone) to 106 cells]. The animals were sacrificed 8-30 weeks after transplantation or if tumors developed in the injected knee. The BM was then harvested. Human engraftment was evaluated by flow cytometry using the following anti-human specific antibodies: anti-CD45-APC Cyanine7 (APC-Cy7) (BioLegend #304014 clone H130); anti-CD33-BV510 (BioLegend #366610); anti-CD34-BV421 (BioLegend #343610); anti-CD38-FITC (BD #555459); anti-CD15 APC (BD #551376); anti-CD19 PE-CY7 (BD #557835); Cell viability was obtained using Propidium Iodide - PI (BioLegend #421301). Analyses were performed using FlowJo software v.10 (Treestar, OR, USA), and the expression of markers was assessed and presented as the percentage of stained cells. The threshold for engraftment detection was 0.1% of human CD45+ cells. All flow cytometry analyses were done using an LSR II (BD Bioscience, CA, USA) or FACSariaII (BD Bioscience, CA, USA) device. Engraftment of >5% of hCD45+ cells with the major CD33+ population (>60% of the hCD45+ cells) was classified as overt leukemia; the presence of 0.1-5% of hCD45+ cells or the presence of >5% of hCD45+ cells with <60% of the CD33+ population in the hCD45+ cells was classified as non-overt leukemia; and the presence of <0.1% of human CD45+ (hCD45+) population was considered as non-engraftment. hCD45+ cells were sorted and underwent whole genome amplification (WGA) for further genotyping assays. CD34+ cells were isolated from PB or BM (following mononuclear cell separation) of AML patients by positive selection using anti-human CD34 microbeads. Enriched CD34+ cells were sorted by FACSariaII (BD bioscience, CA, USA) into four subpopulations of HSPCs according to their surface markers and CD33 expression. The sorted fractions were collected either into Iscove's modified Dulbecco's medium (IMDM) +20% fetal bovine serum (FBS) (Biological Industries #02-023-1A) to be further used in the colony forming unit (CFU) assay or into PBS+2%FBS to be used in the xenotransplantation assay. The CFU assay was adjusted to the generation of single cell-derived subclones. HSPCs were seeded in 1% standard methylcellulose medium supplemented with 37% FBS, 1.2% bovine serum albumin (BSA), 1% L-glutamine, 10-6M b-mercaptoethanol, 1 IU/ml of human erythropoietin (EPO), 50 ng/ml of human stem cell factor (SCF), 25 ng/ml of human FLT3 ligand, 10 ng/ml of interleukin 3 (IL-3), 10 ng/ml of interleukin 6 (IL-6), 10 ng/ml of granulocytecolony stimulating factor (G-CSF), 5 ng/ml of granulocyte-macrophage colony-stimulating factor (GM-CSF), 10 ng/ml of human thrombopoietin (TPO), 100 IU ml penicillin and 100 mg/ml streptomycin. Colonies were counted after 14 days and each of them was picked under sterile conditions. Every individual colony was divided into two cell suspensions to perform both genotyping analysis and xenotransplantation into NSG mice One µl of DNA template was added to a hybridization mix together with an MIP pool (final concentration of 0.05pM per probe) in 1x Ampligase buffer (Epicentre). The mix was incubated in a thermal cycler according to the following program: 98°C for 3 minutes, 85°C for 30 minutes, 60°C for 60 minutes and 56°C for 1 or 2 overnight incubations. The product was mixed with (final concentration in brackets): dNTPs (Larova, 15pM), Betaine (375 mM, Sigma-Aldrich,), NAD+ (1 mM, New England Biolabs), additional Ampligase buffer (0.5x), Ampligase (total of 1.25U, Epicentre) and Phusion HF (0.16U, New England Biolabs). The mix was incubated at 56°C for 60 minutes followed by 72°C for 20 minutes. Enzymatic digestion of linear probes was performed by adding Exonuclease I (4U, New England Biolabs) and Exonuclease III (25U, New England Biolabs). The mix was incubated at 37°C for 2 hours, followed by 80°C for 20 minutes. The final product was amplified using iProof HF Master Mix (Biorad). Samples were pooled and concentrated using AMPure XP beads (Beckman Coulter) at 1.3x volumetric concentration, size-selected (190-370bp) using Blue Pippin (Sage scientific), and sequenced in either NextSeq or Novaseq6000 (Illumina) 2X151bp paired-end run using custom primers as described in the past(Biezuner et al., 2022)

characteristic: description

text: adult

characteristic: developmental stage

text: acute myeloid leukemia

characteristic: individual

text: Homo sapiens

text: Homo sapiens

text: blood

text: FACS Aria

characteristic: protocol

characteristic: sample id

text: Homo sapiens

text: male

characteristic: sex

text: NPM1+ FLT3-ITD+

text: Sample 218


E-MTAB-12238:Sample 219

characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC first public

characteristic: INSDC last update

text: public

text: E-MTAB-12238:Sample 219

text: Repli G

characteristic: WGA

text: ArrayExpress

text: diagnosis

text: acute myeloid leukemia

text: human

text: adult

text: acute myeloid leukemia

characteristic: disease

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: blood

characteristic: organism part

text: FACS Aria

text: Homo sapiens

text: male

text: NPM1+ FLT3-ITD+; CD34 positive

characteristic: subclone

text: Sample 219

characteristic: title

text: 1:10 human CD45 positive out of mouse bone marrow 104


E-MTAB-12238:Sample 22

text: Department of Molecular Cell Biology, Weizmann Institute of Science

text: public

characteristic: INSDC status

characteristic: SRA accession

text: E-MTAB-12238:Sample 22

characteristic: Submitter Id

text: Repli G

text: ArrayExpress

characteristic: broker name

text: diagnosis

characteristic: clinical history

text: acute myeloid leukemia B phenotype

characteristic: clinical information

text: human

characteristic: common name

characteristic: date

characteristic: description

text: adult

characteristic: developmental stage

text: acute myeloid leukemia

text: CD34+, CD38-,CD90+/-, CD45Ra-

characteristic: immunophenotype

characteristic: individual

text: Homo sapiens

text: Homo sapiens

text: bone marrow

text: FACS Aria - H4100 colony

text: Homo sapiens

text: male

text: BCR-ABL/NPM1-/FLT3 ITD-/DNMT3A-

characteristic: time

text: Sample 22


E-MTAB-12238:Sample 220

characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC first public

characteristic: INSDC last update

text: public

text: E-MTAB-12238:Sample 220

text: Repli G

characteristic: WGA

text: ArrayExpress

text: diagnosis

text: acute myeloid leukemia

text: human

text: adult

text: acute myeloid leukemia

characteristic: disease

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: blood

text: FACS Aria

characteristic: protocol

characteristic: sample id

text: Homo sapiens

text: male

characteristic: sex

text: NPM1+ FLT3-ITD+; CD34 positive

text: Sample 220

characteristic: xenograft


E-MTAB-12238:Sample 221

text: public

characteristic: INSDC status

characteristic: SRA accession

text: E-MTAB-12238:Sample 221

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: common lymphoid progenitor

text: diagnosis

characteristic: clinical history

text: acute myeloid leukemia

characteristic: clinical information

text: human

characteristic: common name

characteristic: date

characteristic: description

text: adult

characteristic: developmental stage

text: acute myeloid leukemia

text: CD34+, CD38-,CD90+/-, CD45Ra+

characteristic: immunophenotype

text: Homo sapiens

text: Homo sapiens

text: bone marrow

characteristic: organism part

text: FACS Aria

text: Homo sapiens

text: male

text: NPM1+ FLT3-ITD+

characteristic: subclone

text: Sample 221

characteristic: title


E-MTAB-12238:Sample 222

characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC first public

characteristic: INSDC last update

text: public

text: E-MTAB-12238:Sample 222

characteristic: WGA

text: ArrayExpress

text: common lymphoid progenitor

text: diagnosis

text: acute myeloid leukemia

text: human

text: adult

text: acute myeloid leukemia

characteristic: disease

text: CD34+, CD38-,CD90+/-, CD45Ra+

characteristic: individual

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: bone marrow

text: FACS Aria

characteristic: protocol

characteristic: sample id

text: Homo sapiens

text: male

text: NPM1+ FLT3-ITD+

characteristic: time

text: Sample 222


E-MTAB-12238:Sample 223

text: public

characteristic: INSDC status

characteristic: SRA accession

text: E-MTAB-12238:Sample 223

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: diagnosis

characteristic: clinical history

text: acute myeloid leukemia

characteristic: clinical information

text: human

characteristic: common name

characteristic: date

characteristic: description

text: adult

characteristic: developmental stage

text: acute myeloid leukemia

text: Homo sapiens

text: Homo sapiens

text: bone marrow

characteristic: organism part

text: DNA Purification from BM

text: Homo sapiens

text: male

characteristic: sex

text: NPM1+ FLT3-ITD+

text: Sample 223


E-MTAB-12238:Sample 224

characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC first public

characteristic: INSDC last update

text: public

text: E-MTAB-12238:Sample 224

text: ArrayExpress

text: diagnosis

text: acute myeloid leukemia

text: human

text: adult

text: acute myeloid leukemia

characteristic: disease

characteristic: individual

text: Homo sapiens

text: Homo sapiens

text: bone marrow

text: DNA Purification from BM

text: Homo sapiens

text: male

text: NPM1+ FLT3-ITD+

characteristic: subclone

text: Sample 224

characteristic: title


E-MTAB-12238:Sample 225

text: public

characteristic: INSDC status

characteristic: SRA accession

text: E-MTAB-12238:Sample 225

characteristic: Submitter Id

text: Repli G

characteristic: WGA

text: ArrayExpress

characteristic: broker name

text: common myeloid progenitor

text: relapse

characteristic: clinical history

text: acute myeloid leukemia B phenotype

characteristic: clinical information

text: human

characteristic: common name

text: adult

text: acute myeloid leukemia

characteristic: immunophenotype

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: bone marrow

text: FACS Aria

characteristic: protocol

characteristic: sample id

text: Homo sapiens

text: male

text: BCR-ABL/NPM1-/FLT3 ITD-/DNMT3A-

characteristic: time

text: Sample 225


E-MTAB-12238:Sample 226

characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC first public

characteristic: INSDC last update

text: public

text: E-MTAB-12238:Sample 226

text: Repli G

text: ArrayExpress

text: common myeloid progenitor

text: relapse

text: acute myeloid leukemia B phenotype

text: human

characteristic: date

characteristic: description

text: adult

characteristic: developmental stage

text: acute myeloid leukemia

characteristic: disease

text: Homo sapiens

text: Homo sapiens

text: bone marrow

characteristic: organism part

text: FACS Aria

text: Homo sapiens

text: male

characteristic: sex

text: BCR-ABL/NPM1-/FLT3 ITD-/DNMT3A-

text: Sample 226


E-MTAB-12238:Sample 227

text: public

characteristic: INSDC status

characteristic: SRA accession

text: E-MTAB-12238:Sample 227

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: diagnosis

characteristic: clinical history

text: acute myeloid leukemia

characteristic: clinical information

text: human

characteristic: common name

text: adult

text: acute myeloid leukemia

characteristic: individual

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: bone marrow

text: DNA Purification from BM

text: Homo sapiens

text: female

characteristic: subclone

text: Sample 227

characteristic: title


E-MTAB-12238:Sample 228

characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC first public

characteristic: INSDC last update

text: public

text: E-MTAB-12238:Sample 228

text: ArrayExpress

text: diagnosis

text: acute myeloid leukemia

text: human

characteristic: date

text: adult

text: acute myeloid leukemia

text: Homo sapiens

text: Homo sapiens

text: bone marrow

text: DNA Purification from BM

characteristic: protocol

characteristic: sample id

text: Homo sapiens

text: female

characteristic: time

text: Sample 228


E-MTAB-12238:Sample 229

text: public

characteristic: INSDC status

characteristic: SRA accession

text: E-MTAB-12238:Sample 229

characteristic: Submitter Id

text: Repli G

characteristic: WGA

text: ArrayExpress

characteristic: broker name

text: diagnosis

characteristic: clinical history

text: acute myeloid leukemia

characteristic: clinical information

text: human

characteristic: description

text: adult

characteristic: developmental stage

text: acute myeloid leukemia

characteristic: disease

text: Homo sapiens

text: Homo sapiens

text: blood

characteristic: organism part

text: FACS Aria

text: Homo sapiens

text: male

characteristic: sex

text: NPM1+ FLT3-ITD+; CD45 positive

text: Sample 229

text: human CD45 positive out of mouse bone marrow 79

characteristic: xenograft


E-MTAB-12238:Sample 23

characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC first public

characteristic: INSDC last update

text: public

text: E-MTAB-12238:Sample 23

text: Repli G

text: ArrayExpress

characteristic: cell count

text: relapse

text: acute myeloid leukemia B phenotype

text: human

characteristic: common name

characteristic: date

text: adult

text: acute myeloid leukemia

characteristic: individual

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: blood

text: FACS Aria

text: Homo sapiens

text: male

text: BCR-ABL/NPM1-/FLT3 ITD-/DNMT3A-; CD45 positive

characteristic: subclone

text: Sample 23

characteristic: title

text: human CD45 positive out of mouse bone marrow 15


E-MTAB-12238:Sample 230

text: public

characteristic: INSDC status

characteristic: SRA accession

text: E-MTAB-12238:Sample 230

characteristic: Submitter Id

text: Repli G

characteristic: WGA

text: ArrayExpress

characteristic: broker name

text: diagnosis

characteristic: clinical history

text: acute myeloid leukemia

characteristic: clinical information

text: human

characteristic: description

text: adult

characteristic: developmental stage

text: acute myeloid leukemia

characteristic: disease

text: Homo sapiens

text: Homo sapiens

text: blood

characteristic: organism part

text: FACS Aria

characteristic: protocol

characteristic: sample id

text: Homo sapiens

text: male

text: NPM1+ FLT3-ITD+; CD45 positive

characteristic: time

text: Sample 230


E-MTAB-12238:Sample 231

characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC first public

characteristic: INSDC last update

text: public

text: E-MTAB-12238:Sample 231

text: Repli G

text: ArrayExpress

text: diagnosis

text: acute myeloid leukemia

text: human

characteristic: common name

characteristic: date

text: adult

text: acute myeloid leukemia

characteristic: individual

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: blood

text: FACS Aria

text: Homo sapiens

text: male

characteristic: sex

text: NPM1+ FLT3-ITD+; CD45 positive

text: Sample 231

text: human CD45 positive out of mouse bone marrow 108

characteristic: xenograft


E-MTAB-12238:Sample 232

text: public

characteristic: INSDC status

characteristic: SRA accession

text: E-MTAB-12238:Sample 232

characteristic: Submitter Id

text: Repli G

characteristic: WGA

text: ArrayExpress

characteristic: broker name

text: diagnosis

characteristic: clinical history

text: acute myeloid leukemia

characteristic: clinical information

text: human

characteristic: description

text: adult

characteristic: developmental stage

text: acute myeloid leukemia

characteristic: disease

text: Homo sapiens

text: Homo sapiens

text: blood

characteristic: organism part

text: FACS Aria

text: Homo sapiens

text: male

text: NPM1+ FLT3-ITD+; CD45 positive

characteristic: subclone

text: Sample 232

characteristic: title


E-MTAB-12238:Sample 233

characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC first public

characteristic: INSDC last update

text: public

text: E-MTAB-12238:Sample 233

text: ArrayExpress

text: common lymphoid progenitor

text: diagnosis

text: acute myeloid leukemia

text: human

characteristic: common name

characteristic: date

text: adult

text: acute myeloid leukemia

text: CD34+, CD38-,CD90+/-, CD45Ra+

characteristic: immunophenotype

characteristic: individual

text: Homo sapiens

text: Homo sapiens

text: bone marrow

text: FACS Aria

characteristic: protocol

characteristic: sample id

text: Homo sapiens

text: male

text: NPM1+ FLT3-ITD+

characteristic: time

text: Sample 233


E-MTAB-12238:Sample 234

text: public

characteristic: INSDC status

characteristic: SRA accession

text: E-MTAB-12238:Sample 234

characteristic: Submitter Id

characteristic: WGA

text: ArrayExpress

characteristic: broker name

text: common lymphoid progenitor

text: diagnosis

characteristic: clinical history

text: acute myeloid leukemia

characteristic: clinical information

text: human

characteristic: description

text: adult

characteristic: developmental stage

text: acute myeloid leukemia

characteristic: disease

text: CD34+, CD38-,CD90+/-, CD45Ra+

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: bone marrow

characteristic: organism part

text: FACS Aria

text: Homo sapiens

text: male

characteristic: sex

text: NPM1+ FLT3-ITD+

text: Sample 234


E-MTAB-12238:Sample 235

characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC first public

characteristic: INSDC last update

text: public

text: E-MTAB-12238:Sample 235

text: ArrayExpress

text: relapse

text: acute myeloid leukemia

text: human

characteristic: common name

characteristic: date

text: adult

text: acute myeloid leukemia

characteristic: individual

text: Homo sapiens

text: Homo sapiens

text: bone marrow

text: DNA Purification from BM

text: Homo sapiens

text: female

characteristic: subclone

text: Sample 235

characteristic: title


E-MTAB-12238:Sample 236

text: public

characteristic: INSDC status

characteristic: SRA accession

text: E-MTAB-12238:Sample 236

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: relapse

characteristic: clinical history

text: acute myeloid leukemia

characteristic: clinical information

text: human

characteristic: description

text: adult

characteristic: developmental stage

text: acute myeloid leukemia

text: Homo sapiens

text: Homo sapiens

text: bone marrow

text: DNA Purification from BM

characteristic: protocol

characteristic: sample id

text: Homo sapiens

text: female

text: Sample 236


E-MTAB-12238:Sample 237

characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC first public

text: public

text: E-MTAB-12238:Sample 237

text: Repli G

characteristic: WGA

text: ArrayExpress

text: common lymphoid progenitor

text: diagnosis

text: acute myeloid leukemia

text: human

text: adult

text: acute myeloid leukemia

characteristic: disease

text: CD34+, CD38-,CD90+/-, CD45Ra+

characteristic: immunophenotype

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: blood

characteristic: organism part

text: FACS Aria

text: Homo sapiens

text: male

characteristic: sex

text: NPM1+ FLT3-ITD+

characteristic: time

text: Sample 237

text: 1:10 human CD45 positive out of mouse bone marrow 79

characteristic: xenograft


E-MTAB-12238:Sample 238

characteristic: INSDC last update

text: public

characteristic: INSDC status

characteristic: SRA accession

text: E-MTAB-12238:Sample 238

characteristic: Submitter Id

text: Repli G

text: ArrayExpress

characteristic: broker name

text: common lymphoid progenitor

text: diagnosis

characteristic: clinical history

text: acute myeloid leukemia

characteristic: clinical information

text: human

characteristic: common name

characteristic: date

characteristic: description

text: adult

characteristic: developmental stage

text: acute myeloid leukemia

text: CD34+, CD38-,CD90+/-, CD45Ra+

characteristic: individual

text: Homo sapiens

text: Homo sapiens

text: blood

text: FACS Aria

text: Homo sapiens

text: male

text: NPM1+ FLT3-ITD+

characteristic: subclone

text: Sample 238

characteristic: title


E-MTAB-12238:Sample 239

characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC first public

text: public

text: E-MTAB-12238:Sample 239

text: Repli G

characteristic: WGA

text: ArrayExpress

text: diagnosis

text: acute myeloid leukemia

text: human

text: adult

text: acute myeloid leukemia

characteristic: disease

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: blood

characteristic: organism part

text: FACS Aria

characteristic: protocol

characteristic: sample id

text: Homo sapiens

text: male

characteristic: sex

text: NPM1+ FLT3-ITD+; CD34 positive

text: Sample 239

text: 1:10 human CD45 positive out of mouse bone marrow 108


E-MTAB-12238:Sample 24

characteristic: INSDC last update

text: public

characteristic: INSDC status

characteristic: SRA accession

text: E-MTAB-12238:Sample 24

characteristic: Submitter Id

text: Repli G

text: ArrayExpress

characteristic: broker name

characteristic: cell count

text: relapse

characteristic: clinical history

text: acute myeloid leukemia B phenotype

characteristic: clinical information

text: human

characteristic: common name

characteristic: date

characteristic: description

text: adult

characteristic: developmental stage

text: acute myeloid leukemia

text: Homo sapiens

text: Homo sapiens

text: blood

text: FACS Aria

text: Homo sapiens

text: male

characteristic: time

text: Sample 24

characteristic: xenograft


E-MTAB-12238:Sample 240

characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC first public

text: public

text: E-MTAB-12238:Sample 240

text: Repli G

characteristic: WGA

text: ArrayExpress

text: diagnosis

text: acute myeloid leukemia

text: human

text: adult

text: acute myeloid leukemia

characteristic: disease

characteristic: individual

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: blood

characteristic: organism part

text: FACS Aria

characteristic: protocol

text: Homo sapiens

text: male

text: NPM1+ FLT3-ITD+; CD34 positive

characteristic: subclone

text: Sample 240

characteristic: title


E-MTAB-12238:Sample 241

characteristic: INSDC last update

text: public

characteristic: INSDC status

characteristic: SRA accession

text: E-MTAB-12238:Sample 241

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: common lymphoid progenitor

text: diagnosis

characteristic: clinical history

text: acute myeloid leukemia

characteristic: clinical information

text: human

characteristic: common name

characteristic: date

characteristic: description

text: adult

characteristic: developmental stage

text: acute myeloid leukemia

text: CD34+, CD38-,CD90+/-, CD45Ra+

characteristic: immunophenotype

text: Homo sapiens

text: Homo sapiens

text: bone marrow

text: FACS Aria

characteristic: sample id

text: Homo sapiens

text: male

characteristic: sex

text: NPM1+ FLT3-ITD+

text: Sample 241


E-MTAB-12238:Sample 242

characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC first public

text: public

text: E-MTAB-12238:Sample 242

characteristic: WGA

text: ArrayExpress

text: common lymphoid progenitor

text: diagnosis

text: acute myeloid leukemia

text: human

text: adult

text: acute myeloid leukemia

characteristic: disease

text: CD34+, CD38-,CD90+/-, CD45Ra+

characteristic: individual

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: bone marrow

characteristic: organism part

text: FACS Aria

characteristic: protocol

text: Homo sapiens

text: male

text: NPM1+ FLT3-ITD+

characteristic: time

text: Sample 242


E-MTAB-12238:Sample 243

characteristic: INSDC last update

text: public

characteristic: INSDC status

characteristic: SRA accession

text: E-MTAB-12238:Sample 243

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: relapse

characteristic: clinical history

text: acute myeloid leukemia

characteristic: clinical information

text: human

characteristic: common name

characteristic: date

characteristic: description

text: adult

characteristic: developmental stage

text: acute myeloid leukemia

text: Homo sapiens

text: Homo sapiens

text: bone marrow

text: DNA Purification from BM

text: Homo sapiens

text: female

characteristic: subclone

text: Sample 243

characteristic: title


E-MTAB-12238:Sample 244

characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC first public

text: public

text: E-MTAB-12238:Sample 244

text: ArrayExpress

text: relapse

text: acute myeloid leukemia

text: human

text: adult

text: acute myeloid leukemia

characteristic: disease

characteristic: individual

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: bone marrow

text: DNA Purification from BM

characteristic: sample id

text: Homo sapiens

text: female

characteristic: sex

text: Sample 244


E-MTAB-12238:Sample 245

characteristic: INSDC last update

text: public

characteristic: INSDC status

characteristic: SRA accession

text: E-MTAB-12238:Sample 245

characteristic: Submitter Id

text: Repli G

characteristic: WGA

text: ArrayExpress

characteristic: broker name

text: diagnosis

characteristic: clinical history

text: acute myeloid leukemia

characteristic: clinical information

text: human

characteristic: common name

text: adult

text: acute myeloid leukemia

text: Homo sapiens

text: Homo sapiens

text: blood

characteristic: organism part

text: FACS Aria

characteristic: protocol

text: Homo sapiens

text: male

text: NPM1+ FLT3-ITD+; CD45 positive

characteristic: time

text: Sample 245

text: human CD45 positive out of mouse bone marrow 81

characteristic: xenograft


E-MTAB-12238:Sample 246

characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC first public

text: public

text: E-MTAB-12238:Sample 246

text: Repli G

text: ArrayExpress

text: diagnosis

text: acute myeloid leukemia

text: human

characteristic: date

characteristic: description

text: adult

characteristic: developmental stage

text: acute myeloid leukemia

characteristic: disease

characteristic: individual

text: Homo sapiens

text: Homo sapiens

text: blood

text: FACS Aria

text: Homo sapiens

text: male

text: NPM1+ FLT3-ITD+; CD45 positive

characteristic: subclone

text: Sample 246

characteristic: title


E-MTAB-12238:Sample 247

characteristic: INSDC last update

text: public

characteristic: INSDC status

characteristic: SRA accession

text: E-MTAB-12238:Sample 247

characteristic: Submitter Id

text: Repli G

characteristic: WGA

text: ArrayExpress

characteristic: broker name

text: immature B cell

text: diagnosis

characteristic: clinical history

text: acute myeloid leukemia B phenotype

characteristic: clinical information

text: human

characteristic: common name

text: adult

text: acute myeloid leukemia

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: bone marrow

characteristic: organism part

text: FACS Aria

characteristic: sample id

text: Homo sapiens

text: male

characteristic: sex

text: BCR-ABL/NPM1-/FLT3 ITD-/DNMT3A-

text: Sample 247


E-MTAB-12238:Sample 248

characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC first public

text: public

text: E-MTAB-12238:Sample 248

text: Repli G

text: ArrayExpress

text: immature B cell

text: diagnosis

text: acute myeloid leukemia B phenotype

text: human

characteristic: date

characteristic: description

text: adult

characteristic: developmental stage

text: acute myeloid leukemia

characteristic: disease

text: Homo sapiens

text: Homo sapiens

text: bone marrow

text: FACS Aria

characteristic: protocol

text: Homo sapiens

text: male

text: BCR-ABL/NPM1-/FLT3 ITD-/DNMT3A-

characteristic: time

text: Sample 248


E-MTAB-12238:Sample 249

characteristic: INSDC last update

text: public

characteristic: INSDC status

characteristic: SRA accession

text: E-MTAB-12238:Sample 249

characteristic: Submitter Id

text: Repli G

characteristic: WGA

text: ArrayExpress

characteristic: broker name

characteristic: cell count

text: diagnosis

characteristic: clinical history

text: acute myeloid leukemia

characteristic: clinical information

text: human

characteristic: common name

text: adult

text: acute myeloid leukemia

characteristic: individual

text: Homo sapiens

text: Homo sapiens

text: blood

text: FACS Aria

text: Homo sapiens

text: male

text: NPM1+ FLT3-ITD+; CD45 positive

characteristic: subclone

text: Sample 249

characteristic: title

text: human CD45 positive out of mouse bone marrow 113

characteristic: xenograft


E-MTAB-12238:Sample 25

characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC first public

text: public

text: E-MTAB-12238:Sample 25

text: Repli G

text: ArrayExpress

text: granulocyte monocyte progenitor cell

text: relapse

text: acute myeloid leukemia B phenotype

text: human

characteristic: date

characteristic: description

text: adult

characteristic: developmental stage

text: acute myeloid leukemia

characteristic: disease

characteristic: immunophenotype

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: bone marrow

characteristic: organism part

text: FACS Aria

characteristic: sample id

text: Homo sapiens

text: male

characteristic: sex

text: BCR-ABL/NPM1-/FLT3 ITD-/DNMT3A-

text: Sample 25


E-MTAB-12238:Sample 250

characteristic: INSDC last update

text: public

characteristic: INSDC status

characteristic: SRA accession

text: E-MTAB-12238:Sample 250

characteristic: Submitter Id

text: Repli G

characteristic: WGA

text: ArrayExpress

characteristic: broker name

characteristic: cell count

text: diagnosis

characteristic: clinical history

text: acute myeloid leukemia

characteristic: clinical information

text: human

characteristic: common name

text: adult

text: acute myeloid leukemia

characteristic: individual

text: Homo sapiens

text: Homo sapiens

text: blood

text: FACS Aria

characteristic: protocol

text: Homo sapiens

text: male

text: NPM1+ FLT3-ITD+; CD45 positive

characteristic: subclone

characteristic: time

text: Sample 250

characteristic: title


E-MTAB-12238:Sample 251

characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC first public

text: public

text: E-MTAB-12238:Sample 251

text: ArrayExpress

text: diagnosis

text: acute myeloid leukemia

text: human

characteristic: date

characteristic: description

text: adult

characteristic: developmental stage

text: acute myeloid leukemia

characteristic: disease

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: bone marrow

characteristic: organism part

text: FACS Aria

characteristic: sample id

text: Homo sapiens

text: male

text: NPM1+ FLT3-ITD+; CD34 positive ; CD34 positive

text: Sample 251


E-MTAB-12238:Sample 252

characteristic: INSDC last update

text: public

characteristic: INSDC status

characteristic: SRA accession

text: E-MTAB-12238:Sample 252

characteristic: Submitter Id

characteristic: WGA

text: ArrayExpress

characteristic: broker name

text: diagnosis

characteristic: clinical history

text: acute myeloid leukemia

characteristic: clinical information

text: human

characteristic: common name

text: adult

text: acute myeloid leukemia

characteristic: individual

text: Homo sapiens

text: Homo sapiens

text: bone marrow

text: FACS Aria

characteristic: protocol

text: Homo sapiens

text: male

characteristic: sex

text: Sample 252


E-MTAB-12238:Sample 253

characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC first public

text: public

text: E-MTAB-12238:Sample 253

text: Repli G

text: ArrayExpress

text: diagnosis

text: acute myeloid leukemia

text: human

characteristic: date

characteristic: description

text: adult

characteristic: developmental stage

text: acute myeloid leukemia

text: Homo sapiens

text: Homo sapiens

text: blood

text: FACS Aria

text: Homo sapiens

text: female

text: IDH1(Rx); CD45 positive

characteristic: subclone

characteristic: time

text: Sample 253

characteristic: title

text: human CD45 positive out of mouse bone marrow 120

characteristic: xenograft


E-MTAB-12238:Sample 254

characteristic: INSDC last update

text: public

characteristic: INSDC status

characteristic: SRA accession

text: E-MTAB-12238:Sample 254

characteristic: Submitter Id

text: Repli G

characteristic: WGA

text: ArrayExpress

characteristic: broker name

text: diagnosis

characteristic: clinical history

text: acute myeloid leukemia

characteristic: clinical information

text: human

characteristic: common name

text: adult

text: acute myeloid leukemia

characteristic: disease

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: blood

characteristic: organism part

text: FACS Aria

characteristic: sample id

text: Homo sapiens

text: female

text: IDH1(Rx); CD45 positive

text: Sample 254


E-MTAB-12238:Sample 255

characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC first public

text: public

text: E-MTAB-12238:Sample 255

text: Repli G

text: ArrayExpress

text: B cell

text: diagnosis

text: acute myeloid leukemia

text: human

characteristic: date

characteristic: description

text: adult

characteristic: developmental stage

text: acute myeloid leukemia

characteristic: individual

text: Homo sapiens

text: Homo sapiens

text: blood

text: FACS Aria

characteristic: protocol

text: Homo sapiens

text: male

characteristic: sex

text: NPM1+ FLT3-ITD+

characteristic: subclone

text: Sample 255

text: 1:10 human CD45 positive out of mouse bone marrow 81


E-MTAB-12238:Sample 256

characteristic: INSDC last update

text: public

characteristic: INSDC status

characteristic: SRA accession

text: E-MTAB-12238:Sample 256

characteristic: Submitter Id

text: Repli G

characteristic: WGA

text: ArrayExpress

characteristic: broker name

text: B cell

text: diagnosis

characteristic: clinical history

text: acute myeloid leukemia

characteristic: clinical information

text: human

characteristic: common name

text: adult

text: acute myeloid leukemia

characteristic: disease

text: Homo sapiens

text: Homo sapiens

text: blood

text: FACS Aria

text: Homo sapiens

text: male

text: NPM1+ FLT3-ITD+

characteristic: time

text: Sample 256

characteristic: title

characteristic: xenograft


E-MTAB-12238:Sample 257

characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC first public

text: public

text: E-MTAB-12238:Sample 257

text: Repli G

text: ArrayExpress

characteristic: cell count

text: common lymphoid progenitor

text: diagnosis

text: acute myeloid leukemia

text: human

characteristic: date

characteristic: description

text: adult

characteristic: developmental stage

text: acute myeloid leukemia

text: CD34+, CD38-,CD90+/-, CD45Ra+

characteristic: immunophenotype

characteristic: individual

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: blood

characteristic: organism part

text: FACS Aria

characteristic: sample id

text: Homo sapiens

text: male

text: NPM1+ FLT3-ITD+

text: Sample 257

text: 1:10 human CD45 positive out of mouse bone marrow 113


E-MTAB-12238:Sample 258

characteristic: INSDC last update

text: public

characteristic: INSDC status

characteristic: SRA accession

text: E-MTAB-12238:Sample 258

characteristic: Submitter Id

text: Repli G

characteristic: WGA

text: ArrayExpress

characteristic: broker name

text: common lymphoid progenitor

text: diagnosis

characteristic: clinical history

text: acute myeloid leukemia

characteristic: clinical information

text: human

characteristic: common name

text: adult

text: acute myeloid leukemia

characteristic: disease

text: CD34+, CD38-,CD90+/-, CD45Ra+

text: Homo sapiens

text: Homo sapiens

text: blood

text: FACS Aria

characteristic: protocol

text: Homo sapiens

text: male

characteristic: sex

text: NPM1+ FLT3-ITD+

characteristic: subclone

characteristic: time

text: Sample 258

characteristic: title


E-MTAB-12238:Sample 259

characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC first public

text: public

text: E-MTAB-12238:Sample 259

text: ArrayExpress

text: diagnosis

text: acute myeloid leukemia

text: human

characteristic: date

characteristic: description

text: adult

characteristic: developmental stage

text: acute myeloid leukemia

characteristic: individual

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: bone marrow

characteristic: organism part

text: FACS Aria

text: Homo sapiens

text: male

text: Sample 259


E-MTAB-12238:Sample 26

characteristic: INSDC last update

text: public

characteristic: INSDC status

characteristic: SRA accession

text: E-MTAB-12238:Sample 26

characteristic: Submitter Id

text: Repli G

characteristic: WGA

text: ArrayExpress

characteristic: broker name

text: granulocyte monocyte progenitor cell

text: relapse

characteristic: clinical history

text: acute myeloid leukemia B phenotype

characteristic: clinical information

text: human

characteristic: common name

text: adult

text: acute myeloid leukemia

characteristic: disease

text: CD34+, CD38+, CD10-, CD7-, CD135(FLT3)+, CD45Ra+

characteristic: immunophenotype

text: Homo sapiens

text: Homo sapiens

text: bone marrow

text: FACS Aria

characteristic: protocol

characteristic: sample id

text: Homo sapiens

text: male

text: BCR-ABL/NPM1-/FLT3 ITD-/DNMT3A-

text: Sample 26


E-MTAB-12238:Sample 260

characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC first public

text: public

text: E-MTAB-12238:Sample 260

text: ArrayExpress

text: diagnosis

text: acute myeloid leukemia

text: human

characteristic: date

characteristic: description

text: adult

characteristic: developmental stage

text: acute myeloid leukemia

text: Homo sapiens

text: Homo sapiens

text: bone marrow

text: FACS Aria

text: Homo sapiens

text: male

characteristic: sex

characteristic: subclone

characteristic: time

text: Sample 260

characteristic: title


E-MTAB-12238:Sample 261

characteristic: INSDC last update

text: public

characteristic: INSDC status

characteristic: SRA accession

text: E-MTAB-12238:Sample 261

characteristic: Submitter Id

text: Repli G

characteristic: WGA

text: ArrayExpress

characteristic: broker name

text: hematopoietic multipotent progenitor cell

text: diagnosis

characteristic: clinical history

text: acute myeloid leukemia

characteristic: clinical information

text: human

characteristic: common name

text: adult

text: acute myeloid leukemia

characteristic: disease

text: CD34+, CD38-,CD90+/-, CD45Ra-

characteristic: individual

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: blood

characteristic: organism part

text: FACS Aria

text: Homo sapiens

text: female

text: Sample 261

text: 1:10 human CD45 positive out of mouse bone marrow 120

characteristic: xenograft


E-MTAB-12238:Sample 262

characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC first public

text: public

text: E-MTAB-12238:Sample 262

text: Repli G

text: ArrayExpress

text: diagnosis

text: acute myeloid leukemia

text: human

characteristic: date

text: Protocols: CD34+ cells were isolated from PB or BM (following mononuclear cell separation) of AML patients by positive selection using anti-human CD34 microbeads. Enriched CD34+ cells were sorted by FACSariaII (BD bioscience, CA, USA) into four subpopulations of HSPCs according to their surface markers and CD33 expression. The sorted fractions were collected either into Iscove's modified Dulbecco's medium (IMDM) +20% fetal bovine serum (FBS) (Biological Industries #02-023-1A) to be further used in the colony forming unit (CFU) assay or into PBS+2%FBS to be used in the xenotransplantation assay. The CFU assay was adjusted to the generation of single cell-derived subclones. HSPCs were seeded in 1% standard methylcellulose medium supplemented with 37% FBS, 1.2% bovine serum albumin (BSA), 1% L-glutamine, 10-6M b-mercaptoethanol, 1 IU/ml of human erythropoietin (EPO), 50 ng/ml of human stem cell factor (SCF), 25 ng/ml of human FLT3 ligand, 10 ng/ml of interleukin 3 (IL-3), 10 ng/ml of interleukin 6 (IL-6), 10 ng/ml of granulocytecolony stimulating factor (G-CSF), 5 ng/ml of granulocyte-macrophage colony-stimulating factor (GM-CSF), 10 ng/ml of human thrombopoietin (TPO), 100 IU ml penicillin and 100 mg/ml streptomycin. Colonies were counted after 14 days and each of them was picked under sterile conditions. Every individual colony was divided into two cell suspensions to perform both genotyping analysis and xenotransplantation into NSG mice. xenograft protocol: NOD.Cg-PrkdcscidIl2rgtm1Wjl/SzJ (NSG) mice (The Jackson Laboratory, Bar Harbor, Maine, USA) were maintained and bred under specific pathogen-free conditions at the Biomedical Core Facility (Faculty of Medicine, the Technion, Haifa, Israel). Twenty-four hours prior to transplantation, 8-12-week-old female NSG mice were sub-lethally irradiated with 225 cGy. Primary human cells were intra-femorally injected. The total number of cells transplanted per mouse depended on the number of viable cells obtained from the BM/PB of a patient [ranged from 1 colony (a single cell-derived subclone) to 106 cells]. The animals were sacrificed 8-30 weeks after transplantation or if tumors developed in the injected knee. The BM was then harvested. Human engraftment was evaluated by flow cytometry using the following anti-human specific antibodies: anti-CD45-APC Cyanine7 (APC-Cy7) (BioLegend #304014 clone H130); anti-CD33-BV510 (BioLegend #366610); anti-CD34-BV421 (BioLegend #343610); anti-CD38-FITC (BD #555459); anti-CD15 APC (BD #551376); anti-CD19 PE-CY7 (BD #557835); Cell viability was obtained using Propidium Iodide - PI (BioLegend #421301). Analyses were performed using FlowJo software v.10 (Treestar, OR, USA), and the expression of markers was assessed and presented as the percentage of stained cells. The threshold for engraftment detection was 0.1% of human CD45+ cells. All flow cytometry analyses were done using an LSR II (BD Bioscience, CA, USA) or FACSariaII (BD Bioscience, CA, USA) device. Engraftment of >5% of hCD45+ cells with the major CD33+ population (>60% of the hCD45+ cells) was classified as overt leukemia; the presence of 0.1-5% of hCD45+ cells or the presence of >5% of hCD45+ cells with <60% of the CD33+ population in the hCD45+ cells was classified as non-overt leukemia; and the presence of <0.1% of human CD45+ (hCD45+) population was considered as non-engraftment. hCD45+ cells were sorted and underwent whole genome amplification (WGA) for further genotyping assays. CD34+ cells were isolated from PB or BM (following mononuclear cell separation) of AML patients by positive selection using anti-human CD34 microbeads. Enriched CD34+ cells were sorted by FACSariaII (BD bioscience, CA, USA) into four subpopulations of HSPCs according to their surface markers and CD33 expression. The sorted fractions were collected either into Iscove's modified Dulbecco's medium (IMDM) +20% fetal bovine serum (FBS) (Biological Industries #02-023-1A) to be further used in the colony forming unit (CFU) assay or into PBS+2%FBS to be used in the xenotransplantation assay. The CFU assay was adjusted to the generation of single cell-derived subclones. HSPCs were seeded in 1% standard methylcellulose medium supplemented with 37% FBS, 1.2% bovine serum albumin (BSA), 1% L-glutamine, 10-6M b-mercaptoethanol, 1 IU/ml of human erythropoietin (EPO), 50 ng/ml of human stem cell factor (SCF), 25 ng/ml of human FLT3 ligand, 10 ng/ml of interleukin 3 (IL-3), 10 ng/ml of interleukin 6 (IL-6), 10 ng/ml of granulocytecolony stimulating factor (G-CSF), 5 ng/ml of granulocyte-macrophage colony-stimulating factor (GM-CSF), 10 ng/ml of human thrombopoietin (TPO), 100 IU ml penicillin and 100 mg/ml streptomycin. Colonies were counted after 14 days and each of them was picked under sterile conditions. Every individual colony was divided into two cell suspensions to perform both genotyping analysis and xenotransplantation into NSG mice One µl of DNA template was added to a hybridization mix together with an MIP pool (final concentration of 0.05pM per probe) in 1x Ampligase buffer (Epicentre). The mix was incubated in a thermal cycler according to the following program: 98°C for 3 minutes, 85°C for 30 minutes, 60°C for 60 minutes and 56°C for 1 or 2 overnight incubations. The product was mixed with (final concentration in brackets): dNTPs (Larova, 15pM), Betaine (375 mM, Sigma-Aldrich,), NAD+ (1 mM, New England Biolabs), additional Ampligase buffer (0.5x), Ampligase (total of 1.25U, Epicentre) and Phusion HF (0.16U, New England Biolabs). The mix was incubated at 56°C for 60 minutes followed by 72°C for 20 minutes. Enzymatic digestion of linear probes was performed by adding Exonuclease I (4U, New England Biolabs) and Exonuclease III (25U, New England Biolabs). The mix was incubated at 37°C for 2 hours, followed by 80°C for 20 minutes. The final product was amplified using iProof HF Master Mix (Biorad). Samples were pooled and concentrated using AMPure XP beads (Beckman Coulter) at 1.3x volumetric concentration, size-selected (190-370bp) using Blue Pippin (Sage scientific), and sequenced in either NextSeq or Novaseq6000 (Illumina) 2X151bp paired-end run using custom primers as described in the past(Biezuner et al., 2022)

characteristic: description

text: adult

characteristic: developmental stage

text: acute myeloid leukemia

text: CD34+, CD38-,CD90+/-, CD45Ra-

characteristic: immunophenotype

text: Homo sapiens

text: Homo sapiens

text: blood

text: FACS Aria

characteristic: protocol

characteristic: sample id

text: Homo sapiens

text: female

characteristic: sex

characteristic: subclone

text: Sample 262