title: GSM6705712: mouse liver leukocyte, dasatinib treatment, scRNAseq; Mus musculus; RNA-Seq
library_construction_protocol: Murine liver was dissected following a 20ml PBS flush, and mechanically processed through a 100micron stainless steel sieve with ice-cold RPMI 1640 (with L-glutamine, supplemented with 2% FCS). The tissue slurry was washed, and debris removed through centrifugation with isotonic Percoll. The cell pellet was washed once more and then resuspended in red cell lysis buffer. After another wash, the isolated liver leukocytes are subjected to dasatinib treatment (if applicable), and stained with an antibody cocktail and subsequent viability dye. The treated and untreated samples were sorted on a BD Influx for CD8+ populations of interest (CD44- naive, CD44+PD1-, and CD44+ PD1high) and then washed for Rhapsody antibody-surface sample tag staining and capture following the manufacturer's protocol ("Single Cell Labelling with the BD Single-Cell Multiplexing Kit and BD AbSeq Ab-Oligos" Doc ID: 214419 Rev. 2.0, "Single Cell Capture and cDNA Synthesis with the BD Rhapsody Single-Cell Analysis System" Doc ID: 210966 Rev. 1.0). Library preparation was performed following the protocol "mRNA Whole Transcriptome Analysis (WTA) and Sample Tag Library Preparation Protocol" Doc ID: 23-21712-00 from BD.