2025-08-30
ENA first public
ERC000011
ENA-CHECKLIST
SAMEA114337031
External Id
INSERM U955
INSDC center name
2023-09-07T14:13:36Z
INSDC last update
public
INSDC status
ERS16326021
SRA accession
E-MTAB-13334:2022_42
Submitter Id
3
month
age
ArrayExpress
broker name
not collected
collection date
house mouse
common name
Protocols: 20 nerves per mouse were dissected and HBSS was added (Sigma, ref: H6648-500ML) on ice. After mechanical dissociation, nerves were transfered in a mix of enzymes (0,15% Trypsin (Sigma #T9201), 0,04% Hyaluronidase (Bioconcept #LS005474), 0,3% Collagenase type 2 (Bioconcept #LS004174) and DNAse I (25 μg/mL)) in HBSS, during 2 hours in a water bath. After digestion, nerves were tritured with a 1000 pipette and a solution of 20% FBS in RPMI (Gibco, ref: 42401018) was added to stop digestion. Tubes were centrifuged and supernatant removed. Single viable cells were sorted (BD InfluxTM Cell Sorter) using a nuclear labelling DRAQ7TM Far-Red Fluorescent Live-Cell Impermeant DNA Dye (Abcam, ref: ab109202). Cells were collected in 20% FBS-RPMI solution. Cells were centrifuged, the supernatant removed, and myelin depletion was performed by using anti-myelin beads (Miltenyi Biotec, ref: 130-096-733) in a 0,5% of MACS solution (Miltenyi Biotec, ref: 5130-091-376). Cells were collected after the passage through the magnetic separator. Around 25,000 cells were loaded into one channel of the Chromium system using the V3.1 single cell reagent kit (10X Genomics) to generate single-cell GEMs Following capture and lysis, cDNA was synthesized, then amplified by PCR for 12 cycles as per the manufacturer’s protocol (10X Genomics).
description
Prss56cre,R26mt, Nf1flox/flox
genotype
not collected
geographic location (country and/or sea)
animals kept alone
growth condition
not applicable
isolate
Mus musculus
organism
Mus musculus
organism
subcutaneous nerve
organism part
Mus musculus
scientific_name
male
sex
C57BL/6
strain
2022_42
title