2024-06-05
ENA first public
ERC000011
ENA-CHECKLIST
SAMEA114317220
External Id
UOM
INSDC center name
2023-08-31T09:40:37Z
INSDC last update
public
INSDC status
ERS16306333
SRA accession
E-MTAB-13320:Sublibrary PD2
Submitter Id
22 to 32
week
age
ArrayExpress
broker name
fibroblast-like synoviocyte
cell_type
not collected
collection date
house mouse
common name
Protocols: Hind paws were dissected to remove skin and separate joints, then incubated with collagenase IV (10 mg/ml, Sigma C5138) in DMEM containing 10% FBS and 1% penicillin-streptomycin at 37ºC with shaking for 1-1.5 hours. The cell suspension was passed through a 40 micron filter then cells were centrifuged and the pellet washed with PBS. Podoplanin positive cells were isolated using the EasySep Release Mouse PE Positive Selection kit (Stemcell, 17656) and a Podoplanin-PE conjugated antibody (BioLegend 127407), according to the manufacturer's instructions. Eluted cells were immediately fixed using the Parse Cell Fixation kit (Parse Biosciences SB1003). Fixed cell suspensions were counted using a haemocytometer before being flash frozen in liquid nitrogen and stored at -80ºC prior to barcoding and library preparation. Fixed cells were barcoded using the Evercode Whole Transcriptome Mini kit (Parse Biosciences EC-W01010), in accordance with the manufacturer's instructions. Libraries for sequencing were prepared using the Evercode Whole Transcriptome Mini kit (Parse Biosciences EC-W01010), in accordance with the manufacturer's instructions. Library fragment size and quality was assayed using a 4200 TapeStation (Agilent Technologies).
description
adult
developmental stage
Col6a1-cre Bmal1-flox PER2::LUC
genotype
not collected
geographic location (country and/or sea)
6
individual
not applicable
isolate
Mus musculus
organism
Mus musculus
organism
hindlimb joint
organism part
WT Mini V1 evercode kit
protocol
Mus musculus
scientific_name
male
sex
Crossed DBA1 and C57BL6J
strain
Sublibrary PD2
title