characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2023-08-01T00:17:01Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2023-08-01T00:17:01Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA112878054
characteristics.characteristic: External Id
characteristics.text: UOE
characteristics.characteristic: INSDC center name
characteristics.text: 2023-08-01T00:17:01Z
characteristics.characteristic: INSDC first public
characteristics.text: 2023-08-01T00:17:01Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS14884709
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-12833:Sample 3 V(D)J
characteristics.characteristic: Submitter Id
characteristics.text: 66
characteristics.unit: year
characteristics.characteristic: age
characteristics.text: ArrayExpress
characteristics.characteristic: broker name
characteristics.text: B cell
characteristics.characteristic: cell_type
characteristics.text: human
characteristics.characteristic: common name
characteristics.text: Protocols: Synovial tissue was collected from RA patients undergoing arthroplasty. The fresh tissue was dissected followed by digestion for 2 hours at 37°C in 1 mg ml−1 Collagenase 1 (Sigma-Aldrich). Debris was removed and cells isolated by sequentially passing through 100, 70 and 40μm cell strainers (Corning). The isolated cells were then were resuspended in resuspension media (RPMI-1640 (Gibco) supplemented with 2 mM l-glutamine, 100 U/ml penicillin, and 100 μg/ml streptomycin, with 40% foetal calf serum (FCS)) then an equal volume of freezing media (resuspension media supplemented with 30% dimethyl sulfoxide (DMSO, Fisher Reagents)) was added to give final density of 10x106/ml in 15% DMSO. The sample was then frozen to -80°C in a Mr Frosty Freezing Container (ThermoFisher Scientific) then stored in liquid nitrogen until further use. Synovial tissue samples were thawed and resuspended according to the 10X protocol as described above. Cells were washed and then stained in FACS buffer at 4°C for 20 minutes with fluorescently conjugated antibodies. Single CD19+ve DAPI-ve CD3-ve CD14-ve B cells were then FACS sorted with the FACS Aria II and FACSDiva 8.0.1 (BD Biosciences). Immediately after sorting the CD19+ve B cells were processed through the Chromium Single Cell Platform using the Chromium Next GEM Single Cell 5' Kit v2 (10X Genomics, PN- 1000265), the Chromium Single Cell Human BCR Amplification Kit (10X Genomics, PN-1000253), and the Chromium Next GEM Chip K Single Cell Kit (10X Genomics, PN-1000287) as per manufacture’s protocol.
characteristics.characteristic: description
characteristics.text: rheumatoid arthritis
characteristics.characteristic: disease
characteristics.text: 3
characteristics.characteristic: individual
characteristics.text: not applicable
characteristics.characteristic: isolate
characteristics.text: Homo sapiens
characteristics.characteristic: organism
characteristics.text: Homo sapiens
characteristics.characteristic: organism
characteristics.text: synovial tissue
characteristics.characteristic: organism part
characteristics.text: Homo sapiens
characteristics.characteristic: scientific_name
characteristics.text: female
characteristics.characteristic: sex
characteristics.text: Sample 3 V(D)J
characteristics.characteristic: title