2025-08-22
ENA first public
ERC000011
ENA-CHECKLIST
Institute of Molecular Genetics of the Czech Academy of Sciences
INSDC center name
public
INSDC status
ERS22563517
SRA accession
E-MTAB-14686:LK_WT_3m_3
Submitter Id
3
month
age
ArrayExpress
broker name
late hematopoietic progenitors (GMP, CEM, MEP) = LK population
cell_type
not collected
collection date
house mouse
common name
Protocols: Mice were sacrificed by cervical dislocation, femurs and tibias were isolated, and crunched using a pestle and mortar. Sorting of bone marrow hematopoietic stem cells and progenitors was performed by two subsequent steps. First, the Lin+ fraction of the BM cells was labeled using biotinylated lineage markers CD45/B220 (RA3-6B2), CD3 (145-2c11), Ter119 (TER-119), Gr1 (RB6-8C5), and CD11b (M1/70). These cells were then incubated with anti-biotin magnetic beads (Miltenyi Biotec) and were isolated using MACS separator. Second, the Lin- fraction of the BM was labeled with the c-Kit PE (2B8), Sca-1 APC (E13-161.7), CD48 FITC (HM48-1), CD150 Pe-Cy7 (TC15-12F12.2) antibodies and with streptavidin-eFluor450. Cell suspensions were stained with Hoechst 33258 to exclude dead cells and HSCs were sorted using Influx instrument (BD Biosciences). Total RNA was isolated using RNeasy Micro Kit (Qiagen) as described in the manufacturer’s protocol. Libraries were prepared with a SMARTer® Stranded Total RNA-Seq – Pico Input Mammalian library preparation kit v2 (Takara).
description
adult
developmental stage
wild-type
genotype
not collected
geographic location (country and/or sea)
15
individual
not applicable
isolate
Mus musculus
organism
Mus musculus
organism
bone marrow
organism part
Mus musculus
scientific_name
B
sequencing run
male
sex
C57BL/6
strain
LK_WT_3m_3
title