title: FL_iEG
organism: Mus musculus
description: Protocols: E14.5 fetal liver cells from CTRL and iETO2-GLIS2 (iEG) embryos were stained for flow cytometry using standard protocols and the LT-HSC populations were flow sorted according to the following markers : LT-HSC: LIN-Kit+Sca1+CD135-CD34-CD48-CD150+ MPP4: LIN-Kit+Sca1+CD135+CD34+CD48+CD150-. Cell populations were flow sorted, cultured 24 hours in RPMI supplemented with 10% FBS, murine cytokines (IL3, IL6, SCF, TPO, FLT3L) and 100ng/ml of Doxycycline to induce ETO2-GLIS2 expression. 24 hours post-induction, we performed cell lysis, transposition and purification step, the transposed DNA fragments were amplified by polymerase chain reaction (PCR) between 12 and 18 depending on the number of cells at the beginning (50,000 to 6,000) using adapters from the Nextera index kit (Illumina). PCR purification was performed using Agencourt AMPure XP magnetic beads (Beckman Coulter A63880) in order to remove large fragments and remaining primers. Library quality was assessed using an Agilent 2100 Bioanalyzer using a High Sensitivity DNA chip (Agilent Technologies 5067-4626). Libraries were constructed using the Nextera index kit (Illumina).
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2019-10-15T04:03:41Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2019-09-26T14:56:38Z
attributes.tag: External Id
attributes.value: SAMEA5977205
attributes.tag: INSDC center name
attributes.value: INSERM, Paris, France Equipe Labellise Ligue Contre le Cancer Institut Gustave Roussy, Villejuif, France Universite Paris Diderot, Paris, France
attributes.tag: INSDC first public
attributes.value: 2019-10-15T04:03:41Z
attributes.tag: INSDC last update
attributes.value: 2019-09-26T14:56:38Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-8375:FL_iEG
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell type
attributes.value: hematopoietic progenitor cell
attributes.tag: common name
attributes.value: house mouse
attributes.tag: developmental stage
attributes.value: E14.5
attributes.tag: disease
attributes.value: leukemia
attributes.tag: genotype
attributes.value: with fusion ETO2GLIS2
attributes.tag: organism part
attributes.value: liver
attributes.tag: sample name
attributes.value: E-MTAB-8375:FL_iEG
attributes.tag: scientific_name
attributes.value: Mus musculus
attributes.tag: strain
attributes.value: C57/B6