characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2022-06-06T00:13:42Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2022-06-06T00:13:42Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA12132960
characteristics.characteristic: External Id
characteristics.text: Institut Necker Enfants Malades (INEM), INSERM U1151/CNRS UMS 8253, Universite de Paris, Paris, France
characteristics.characteristic: INSDC center name
characteristics.text: 2022-06-06T00:13:42Z
characteristics.characteristic: INSDC first public
characteristics.text: 2022-06-06T00:13:42Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS9779569
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-11294:Sample 3
characteristics.characteristic: Submitter Id
characteristics.text: 14
characteristics.unit: week
characteristics.characteristic: age
characteristics.text: ArrayExpress
characteristics.characteristic: broker name
characteristics.text: immune cell
characteristics.characteristic: cell type
characteristics.text: house mouse
characteristics.characteristic: common name
characteristics.text: Protocols: C56BL/6 mice (10 to 12-week-old) were immunized subcutaneously at the base of the tail with 100 μg of MOG (35-55) emulsified in complete Freund's adjuvant containing 500 μg of Mycobacterium tuberculosis. Pertussis toxin (200 ng; List Biological Laboratories) was injected intravenously at days 0 and 2 post-immunization. At day 17 post immunization, central nervous system CD45-positive immune cells were isolated and loaded onto a single channel of a 10x Chromium chip for each sample. The scRNA-seq libraries were generated using Chromium Next GEM Single Cell V(D)J Reagent Kit v.1.1 with Feature Barcoding (10X Genomics) according to the manufacturer’s protocol. Gene expression (mRNA) libraries were constructed. Briefly, cells were counted and up to 20 000 cells were loaded in the 10X Genomics Chromium Controller to generate single-cell gel-beads in emulsion. After reverse transcription, gel-beads in emulsion were disrupted. Barcoded complementary DNA was isolated and amplified by PCR. Following fragmentation, end repair and A-tailing, sample indexes were added during index PCR. The purified libraries were sequenced on a Novaseq S2 flowcell (Illumina) with 26 cycles of read 1, 8 cycles of i7 index and 91 cycles of read 2, targeting a median depth of 50000 reads per cell for gene expression.
characteristics.characteristic: description
characteristics.text: adult
characteristics.characteristic: developmental stage
characteristics.text: autoimmune encephalomyelitis
characteristics.characteristic: disease
characteristics.text: CD45 positive
characteristics.characteristic: immunophenotype
characteristics.text: Mus musculus
characteristics.characteristic: organism
characteristics.text: Mus musculus
characteristics.characteristic: organism
characteristics.text: spinal cord
characteristics.characteristic: organism part
characteristics.text: Mus musculus
characteristics.characteristic: scientific_name
characteristics.text: female
characteristics.characteristic: sex
characteristics.text: C56BL/6
characteristics.characteristic: strain
characteristics.text: Sample 3
characteristics.characteristic: title