GSM8722242: D7 enriched NK cells_ADT_library2; Mus musculus; OTHER
C57Bl/6 splenic CD8 T cells were enriched by STEMCELL CD8 T cell negative selection kit. All samples were stained with TotalSeq-C DNA Barcoded CD45.1, CD45.2, Thy1.1, Thy1.2 antibodies. Individual mouse were labeled with TotalSeq-C DNA-Barcoded hashtags 1-5 (for adult or infant). Cells were washed and equilabrated and combined. Combined cell pool was then labeled with TotalSeq-C DNA-Barcoded KLRG1, CD127, CD4, CD8a. CITE libraries were constructed using the Chromium Next GEM Single Cell 5' Reagent Kit v2 (10x Genomics) with a 5' Feature Barcode Kit (10x Genomics) for antibody-derived tag (ADT) amplification according to the manufacturer's instructions. The ADT library pool and scRNA-seq library pool were each sequenced on one lane of an S4 flow cell on an Illumina NovaSeq 6000 (~2.5 billion 2x100 bp reads per lane). Quantification of scRNA-seq library pools was performed by pseudoalignment to the human transcriptome (Gencode v24 annotation derived from GRCh38) using kallisto v0.46.2 in “BUS” mode