Mus musculus
Protocols: Single-cell suspensions were prepared from pre-leukemic mouse bone marrow under sterile conditions. Red blood cells were lysed with Red Blood Cell Lysis Solution (Miltenyi Biotec) and washed with staining media [SM: Hanks' Balanced Salt Solution (Thermo Fisher Scientific), 2% fetal bovine serum (FBS, Wisent), 10mM HEPES, pH 7.2]. Depletion of CD11b+ cells was carried out by magnetic cell separation with autoMACS (Miltenyi Biotec) per manufacturer's instructions. Bone marrow cells were stained with fluorochrome-conjugated antibodies for 30 minutes in the dark at room temperature and CD8β- CD19+ CD127+ Sca-1+ were sorted on a MoFlo Astrios (Beckman Coulter) in the Flow Cytometry Facility at the Hospital for Sick Children. Polyinosinic:polycytidylic acid (pIpC) administration to activate Mx1-cre, and consequently Prdm14 expression, was performed at 8 weeks of age with one intraperitoneal injection of 250µg. The 10X Genomics Chromium platform was used for single cell RNA-sequencing (scRNA-seq) at the Princess Margaret Genomics Centre. Determination of the viability and density of sorted cells were carried out by a Countess Automated Cell Counter (ThermoFisher Scientific). Capture and encapsulation of cells (approximately 2,000 per sample) was followed by cDNA amplification and library preparation for sequencing.
ENA-FIRST-PUBLIC
2023-12-01T00:42:47Z
ENA-LAST-UPDATE
2023-12-01T00:42:47Z
External Id
SAMEA114009063
INSDC center name
Genetics and Genome Biology, SickKids Research Institute, Toronto, ON, Canada. Department of Molecular Genetics, University of Toronto, Toronto, ON, Canada
INSDC first public
2023-12-01T00:42:47Z
INSDC last update
2023-12-01T00:42:47Z
INSDC status
public
Submitter Id
E-MTAB-13160:1_GFPpos_BM_Preleuk
age
8
broker name
ArrayExpress
collection date
not collected
common name
house mouse
developmental stage
adult
genotype
R26PR;Mx1-cre
geographic location (country and/or sea)
not collected
immunophenotype
CD11b- CD19+ CD127+ Sca-1+; GFP positive
individual
JCLR_scRNA1
isolate
not applicable
organism part
bone marrow
sample name
E-MTAB-13160:1_GFPpos_BM_Preleuk
scientific_name
Mus musculus
sex
female
strain
C57BL/6J