2024-05-18
ENA first public
ERC000011
ENA-CHECKLIST
SAMEA113565820
External Id
KIS
INSDC center name
2023-05-31T12:02:21Z
INSDC last update
public
INSDC status
ERS15561968
SRA accession
E-MTAB-13036:PK2326_ymito
Submitter Id
2 to 4
month
age
ArrayExpress
broker name
stem cell
cell_type
house mouse
common name
Protocols: Mouse small intestinal crypts were isolated as follows. Small intestines were flushed with ice-cold PBS and opened longitudinally, and mucus was removed by gentle rubbing. The intestine was cut into small fragments and incubated with three changes of 10 mM EDTA in PBS on ice for 1 h and 45 mins. Epithelium was then detached by vigorous shaking. To enrich for crypts, the tissue suspension was filtered through 70-μm nylon mesh. Single cells were isolated by dissociating crypts in TrypLE Express (Gibco) with 1,000 U/ml of DNase I (Roche) at 32 °C for 2 minutes. Cells were washed and stained with the following antibodies: CD31–PerCP-Cy5.5 (BD, Mec13.3), CD45–PerCP-Cy5.5 (BD, 30-F11), Ter119– PerCP-Cy5.5 (BD Ter119), CD326–BV786 (BD, G8.8) and CD24–BV421 (BD, M1/69), all at 1:500 dilution. Finally, cells were resuspended in HBSS containing 0.2% BSA supplemented with 7-AAD (Life) (2 μg/ml) for live-cell separation. Cells were sorted using a FACSAria Fusion (BD Biosciences). RNA extraction was performed using Trizol according to manufacturer’s instructions for RNA isolation. Samples were first treated with HL-dsDNase (Articzymes 80200-050) to remove residual DNA. NuGEN Ovation SoLo RNA-Seq kit was used for Illumina library preparations (NuGEN Technologies).
description
adult
developmental stage
Lgr5 EGFP-CreERT2/+ Rosa26 Snap-Omp25/+
genotype
PK2326
individual
not applicable
isolate
Mus musculus
organism
Mus musculus
organism
epithelium of small intestine
organism part
young mitochondria
phenotype
Mus musculus
scientific_name
male
sex
C57BL/6J
strain
PK2326_ymito
title