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. Cleavage Under Targets and Release Using Nuclease (CUT&RUN) was carried out as previously described (Meers et al., 2019) on sorted cells. 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. Cleaved DNA was extracted with phenol-chloroform with no changes to the default protocol Libraries were generated using the NEBNext® Ultra II DNA Library Prep Kit (New England BioLabs) per manufacturer's instructions.
ENA-FIRST-PUBLIC
2023-12-01T00:42:47Z
ENA-LAST-UPDATE
2023-12-01T00:42:47Z
External Id
SAMEA114028125
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-13159:JCLR_001
age
8
broker name
ArrayExpress
cell_type
bone marrow cell
collection date
not collected
common name
house mouse
developmental stage
adult
genotype
R26PR;Mx1-cre
geographic location (country and/or sea)
not collected
immunophenotype
CD19+ CD127+ Sca-1+
individual
JCLRmouse1
isolate
not applicable
organism part
bone marrow
sample name
E-MTAB-13159:JCLR_001
scientific_name
Mus musculus
sex
female
strain
C57BL6