2024-07-24
ENA first public
ERC000011
ENA-CHECKLIST
SAMEA115031429
External Id
UH
INSDC center name
2023-12-14T16:42:38Z
INSDC last update
public
INSDC status
ERS17478362
SRA accession
E-MTAB-13624_2:sample59
Submitter Id
10
month
age
ArrayExpress
broker name
gastric mucosa cell
cell_type
not collected
collection date
house mouse
common name
Protocols: Antral polyps and adjacent antral mucosa from Twist2Cre:Lkb1fl/+;Tdtomato+/- mice were dissociated into single cells as follows: the tissue was flushed with PBS, minced into < 1 mm pieces with scissors, and placed in a 15 ml falcon tube filled with 5 ml of a collagenase mixed solution (collagenase II (Gibco 17101-019) + IV (ThermoFischer 17104019), 1:1 ratio at 2 mg/ml). The samples were incubated horizontally in a 37°C shaking incubator (220 rpm) for 40 min. During this incubation time, pieces were pipetted up and down 10 times every 10 min, and the collagenase solution was changed after 20 minutes, collecting the supernatant containing the single cells. The supernatant (~10 ml) was then transferred to a clean Falcon tube, and collagenases were inactivated with cold DMEM supplemented with 20% fetal bovine serum (FBS). The samples were passed through a 70 um cell strainer, centrifuged for 10 min at 1100 rpm, and single cells were resuspended in 1–2 ml of PBS. Single-cell suspensions were stained with 7 AAD (LifeTechnologies #A1310) at a 1:1000 dilution prior to FACS sorting to exclude dead cells. 10.000 live cells per sample were sorted at ultra-purity using a Sony SH800Z Cell Sorter. The 10X v3 library kit (Library Construction Kit from 10X Genomics #1000190) was used for scRNA-seq library preparation. The 10X v3 library kit (Library Construction Kit from 10X Genomics #1000190) was used for scRNA-seq library preparation.
description
adult
developmental stage
Twist2Cre;Lkb1fl/+
genotype
not collected
geographic location (country and/or sea)
3315
individual
not applicable
isolate
Mus musculus
organism
Mus musculus
organism
gastric mucosa
organism part
gastric mucosa
sampling site
Mus musculus
scientific_name
female
sex
ICR mixed background
strain
sample59
title