title: GSM7256688: SP_pool_4; Mus musculus; RNA-Seq
library_construction_protocol: Tumous were digested with DNase I (100 µg/mL) and Collagenase IV (200 U/mL) for 40 minutes at 37°C and subsequently mashed through a 70 µm and  a 30 µm cell strainer. Cells were labelled with TotalSeq-C Hashtag antibodies (Hashtags 1-4, Biolegend) and stained for surface markers and viability. Intratumoural CD8+ T cells were then sorted (live CD8a+ CD3+ CD45+ cells). Library construction was carried out by using the following kit and protocol: Chromium Next GEM Single Cell 5' Reagent Kits v2 User Guide with Feature Barcode technology for Cell Surface Protein, Rev D. For sequencing, libraries were pooled fulfilling the minimum criteria: 2x10^4 reads/cell for gene expression libraries and 5x10^3 reads/cell for TCR libraries. Pooled libraries were sent to Novogene for sequencing.