title: GSM9386696: CIA mouse, GEX; Mus musculus; RNA-Seq
library_construction_protocol: Spleens were ground through a 70um mesh filter before being spun down 500g. Followed by a 5-minute incubation on ice in ACK buffer to lyse red blood cells.  Cells were spun down 500g, counted and pooled together before being washed in PBS and resuspended at 1x106 cells/100ul prior to barcoding This enriched cell suspension was processed using the Chromium 5′ Single-Cell Gene Expression system (10X Genomics) to generate scRNA seq libraries following the manufacturer's protocol. Briefly, single-cell suspensions with ≥85% viability were incubated with a pool of oligonucleotide-conjugated antibodies (BioLegend TotalSeq™) for sample indexing. Labeled cells were loaded into a Chromium Next GEM Chip G to generate Gel Bead-In-Emulsions (GEMs), enabling barcoding of individual cells and transcripts. Reverse transcription, cDNA amplification, and separate library construction for gene expression and feature barcodes were performed as described in the 10x Genomics protocol. Libraries were quantified, pooled, and sequenced on an Illumina NovaSeq platform with recommended read configurations.