organism: Mus musculus
description: 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).
attributes.tag: ENA first public
attributes.value: 2025-08-22
attributes.tag: INSDC center name
attributes.value: Institute of Molecular Genetics of the Czech Academy of Sciences
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-14686:HSC_MUT_3m_2
attributes.tag: age
attributes.value: 3
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell_type
attributes.value: hematopoietic stem cell
attributes.tag: collection date
attributes.value: not collected
attributes.tag: common name
attributes.value: house mouse
attributes.tag: developmental stage
attributes.value: adult
attributes.tag: genotype
attributes.value: homozygous Jak2 R1063H
attributes.tag: geographic location (country and/or sea)
attributes.value: not collected
attributes.tag: individual
attributes.value: 6
attributes.tag: isolate
attributes.value: not applicable
attributes.tag: organism part
attributes.value: bone marrow
attributes.tag: sample name
attributes.value: E-MTAB-14686:HSC_MUT_3m_2
attributes.tag: scientific_name
attributes.value: Mus musculus
attributes.tag: sequencing run
attributes.value: A
attributes.tag: sex
attributes.value: female
attributes.tag: strain
attributes.value: C57BL/6