2025-11-28
ENA first public
ERC000011
ENA-CHECKLIST
Roche Pharma Research & Early Development (pRED) Roche Innovation Center Basel
INSDC center name
public
INSDC status
ERS24548695
SRA accession
E-MTAB-15144:E6_Nivo_Rela_d28
Submitter Id
6 to 8
week
age
ArrayExpress
broker name
T cell
cell type
not collected
collection date
house mouse
common name
Protocols: Mice were sacrificed, and tumor tissue was isolated. The tumor tissue was disrupted and then digested in an enzyme mix containing RPMI with 10 mg/ml DNAse and 0.25 mg/ml Liberase for 30 minutes at 37°C. The resulting tissue mix was filtered to obtain a single-cell suspension. Lymphocytes were mechanically isolated from draining lymph nodes, filtered, and resuspended to a single-cell suspension. The resulting single-cell suspensions from tumors and lymph nodes were stored in liquid nitrogen in pZerve freezing medium. After thawing a batch of samples, the cells were stained with a mix of FACS and oligo-labelled antibodies and sorted for CD3+T cells before performing scRNA-seq. Prior to sorting, the cells were washed with PBS, and cell number and viability were evaluated. Approximately 1 million cells per sample were resuspended in PBS and incubated with a Fc blocking reagent. The mix of FACS and oligo-labelled antibodies was added, and after incubation and washing, the cells were resuspended for sorting. The cell number and viability of the sorted cells were determined, and a total of 10,000 viable cells per sample were loaded into the 10x Genomics Chromium Connect Instrument. cDNA and library preparation was performed according to the manufacturer’s indications using the 10x Genomics Chromium Next GEM Automated Single Cell 5’ Reagent Kits v2 with TCR and feature barcoding.
description
adult
developmental stage
Panc02-Fluc
genotype
not collected
geographic location (country and/or sea)
E6
individual
not applicable
isolate
Mus musculus
organism
Mus musculus
organism
pancreas
organism part
Mus musculus
scientific_name
female
sex
C57BL/6J
strain
E6_Nivo_Rela_d28
title