title: GSM6945061: mSTAR1302, scRNAseq; Mus musculus; RNA-Seq
library_construction_protocol: Tissue tumor samples were gently cut to about 1 mm3 pieces placed in gentleMACS C-tubes (Miltenyi). Tissues were digested enzymatically containing 20mM HEPES(Gibco), 1mg/mL Collagenase (Sigma), 0.1mg/mL DNase (Roche) for 30 mins in a 37°C shaker with gentle agitation. The tissue pieces were gently dissociated with mechanical force using the GentleMACS Octo Dissociator (Miltenyi Biotec). Cells were then filtered through a 70μm cell strainer and suspended cells were subsequently washed and centrifuged for 5min at 300g. After tissue digestion and dissociation, CD4+ and CD8+ TILs were isolated using the mouse CD4/CD8 (TIL) MicroBeads (Miltenyi) Samples were loaded onto a 10x Chromium instrument to generate 3' gene expression libraries (10x Genomics), following manufacturer's protocol. Libraries were sequenced on NovaSeq to a minimum sequencing depth of 50,000 reads per cell.