title: GSM9662616: Tet2fl/fl CD45+ TILs, TCR, biol rep 1; Mus musculus; OTHER
library_construction_protocol: Tumor-infiltrating leukocytes were isolated from liver metastatic tumors by enzymatic digestion with collagenase A (0.75 mg/ml) and DNase I (50 µg/ml) followed by density gradient centrifugation using Histopaque-1083. Cells were washed and resuspended in PBS supplemented with 1% BSA, stained with BV421 anti-mouse CD45.2 antibody (clone 104, BioLegend, Cat#109832) and 7-AAD viability solution (ThermoFisher Scientific, Cat#00-6993-50), and CD45+ live cells were sorted using BD FACSAria III (BD Biosciences, RRID: SCR_016695). Sorted CD45+ live cells were processed to generate barcoded 5' gene expression (GEX) libraries and TCR V(D)J-enriched libraries using the Chromium Next GEM Single Cell 5' Kit v1.1 (10x Genomics, Cat#1000165) and GEM Single Cell V(D)J Reagent Kits v1.1 (10x Genomics, Cat#1000181), targeting 10,000 cells per library, following the manufacturer's instructions (CG000208 Rev G). Library quality and concentration were assessed using the Bioanalyzer High Sensitivity DNA Kit (Agilent Technologies, Cat#5067-4626) and KAPA Library Quantification Kit (Kapa Biosystems, Cat#KK4873).