GSM9683365: Multiplexed_5; Mus musculus; OTHER
Mouse splenic and thymic CD8+ T cells were isolated using CD8a+ T cell isolation kit (Miltenyi Biotec, Cat#130-104-075). Mouse splenic naïve CD8+ T cells were isolated using Naive CD8a+ T Cell Isolation Kit, mouse (Miltenyi Biotec, Cat#130-096-543). For isolating CD8+ tumor infiltrating lymphocytes (TILs), tumors were first cut into small pieces of 2-4mm and digested using tumor dissociation kit at 37°C for 45 minutes with gentle agitation (Miltenyi Biotec, Cat#130-096-730). Dead cells were further removed using Dead cell Removal kit (Miltenyi Biotec, Cat# 130-090-101). CD8+ TILs were then isolated using CD8 (TIL) microbeads, mouse (Miltenyi Biotec, Cat#130-116-478). In this TCR sequencing cohort, single cell suspensions were prepared using Evercode TCR kit. cDNA libraries were generated from whole-transcriptome amplification products containing V(D)J segments spanning the CDR3 regions of TCRα (TRA) and TCRβ (TRB) chains, followed by ligation of Illumina-compatible adapters.