GSM9491366: CITE-seq, Day28 ILCs, Gene Expression, First Run; Homo sapiens; OTHER
Live, human CD45+ cells were sorted using fluorescence-activated cell sorting (FACS) to remove dead cells and stromal contamination. Immediately post-sort, cells were stained with TotalSeq-C antibodies, counted and loaded onto the 10x Genomics Chromium Controller to generate Gel Beads-in-emulsion (GEMs). Lysis and barcoded reverse transcription of both mRNA and antibody-derived tags (ADTs) occurred inside the GEMs using the Chromium Next GEM Single Cell 5' Reagent Kit v2 (10x Genomics). Post-GEM-RT cleanup and cDNA amplification were performed according to the 10x Genomics Chromium Next GEM Single Cell 5' v2 User Guide.  Two distinct library types were constructed from the amplified cDNA:  Gene Expression (GEX) Libraries: Generated from the cDNA utilizing the 5' gene expression library construction kit.  Cell Surface Protein (ADT) Libraries: Generated from the supernatant of the cDNA amplification reaction, which contained the TotalSeq-C antibody-derived tags. These were indexed using the appropriate 10x Genomics Dual Index Kit TT Set A.  Final libraries were quantified and pooled for sequencing.