2024-05-28
ENA first public
ERC000011
ENA-CHECKLIST
Institute of Molecular Genetics of the Czech Academy of Sciences
INSDC center name
public
INSDC status
ERS18431543
SRA accession
E-MTAB-13908:786
Submitter Id
12
week
age
11
week
age (competitor)
33
week
age (recipient)
ArrayExpress
broker name
LSK (Lin-SCA1+KIT+) bone marrow cells
cell_type
not collected
collection date
house mouse
common name
Protocols: Donor Wbp1l-/- or WT mice were sacrificed by Cervical dislocation. Femurs, tibias and spines were separated, cleaned from muscle tissue, crushed using mortar and pestle and filtered through nylon mesh. Cell suspension was then reconstituted in ACK buffer (150 mM NH4Cl, 0.1 mM EDTA (disodium salt), 1 mM KHCO3) to remove erythrocytes. Next, the bone marrow cells were stained with mix of lineage antibodies (CD3e, B220, Ter119, Gr-1 and CD11b) and separated on autoMACS separator to enrich for immature populations. The sorted unstained fraction was further stained with Lineage antibody cocktail together with KIT, SCA1, Ly5.1, and Ly5.2 antibodies. Ly5.2+Ly5.1- LSK bone marrow cells (Lin-SCA1+KIT+) were then sorted on BDinflux machine. Sorted cells were mixed 1:1 with WT bone marrow competitor cells (Ly5.2+Ly5.1+, extracted using the same procedure as the donor cells) and the mixtures (4x106 cells per recipient) were transplanted into the lethally irradiated (7Gy) Ly5.1+ recipient mice by injection into the tail vein. 22 weeks after transplantation the recipient mice were sacrificed and the donor Ly5.2+Ly5.1- LSK cells were purified. RNA from purified LSK cells was isolated using Rneasy Micro Kit (Qiagen) and treated with DNAse I (Qiagen) for 15 min. Libraries were prepared with a SMARTer® Stranded Total RNA-Seq – Pico Input Mammalian library preparation kit v2 (Takara).
description
adult
developmental stage
Wbp1l-/- Ly5.2+/+
genotype
Wbp1l+/+ Ly5.1+Ly5.2+
genotype (competitor)
Wbp1l+/+ Ly5.1+/+
genotype (recipient)
not collected
geographic location (country and/or sea)
45L-29690
individual
45L-28727
individual (competitor)
45L-28786
individual (recipient)
not applicable
isolate
Mus musculus
organism
Mus musculus
organism
bone marrow
organism part
Mus musculus
scientific_name
female
sex
C57BL/6J x Wbp1ltm2a(EUCOMM)Hmgu
strain
57BL/6J x C57BL/6NCrl-Ly5.1
strain (competitor)
C57BL/6NCrl-Ly5.1
strain (recipient)
786
title