title: GSM6792030: CD8 T-cells, Feature barcode-derived cDNA (Sc_D3); Mus musculus; OTHER
library_construction_protocol: CD8+ T cells were enriched from splenocytes by negative selection with anti-CD4, anti-CD19, anti-BD220, and anti-TER19 and magnetic streptavidin beads.  Enriched cells were stained for CD8,  Thy1.1 and Thy1.2 to identify donor P14.  Cells were then sorted by FACS, hashed, and analyzed together to avoid batch effects.  Anti-mouse mouse MHC H-2 hashtag antibodies (Biolegend TotalSeq) were used to label the separate FACS-purified CD8 Tcell subsets Cells were washed, counted and pooled to a final concentration of ~1,600 cells/ul.  A total of ~50,000 cells were loaded into one lane of the single cell A chip kit. Library was prepared using Chromium Single Cell 3ʹ v2 protocol (10x Genomics) according to manufacturer recommendations. The final library size distribution were assessed using bioanalyzer analysis and quantified with the NEBNext library quantification kit. The cDNA and HTO libraries were pooled 10:1 and sequenced to a depth of 50,000 reads per cell for the cDNA and 2,000 reads per cell for the HTO library. Libraries were sequenced on the Illumina NovaSeq with the following 10X read format; Read 1 25bp, indexi7 8bp, and Read 2 98bp.