title: GSM9217268: 4T1 Cmas KO, TILs; Mus musculus; RNA-Seq
library_construction_protocol: Tumors were resected at day 15 post injection and immediately dissociated using a tumor dissociation kit and GentleMACS dissociator (both Miltenyi). CD45 positive cells were then isolated with anti-CD45 microbeads (Miltenyi) following a previously described protocol. Cells were counted using NucleoCounter NC250 (Chemometec) following the manufacturer instructions and CD45 positivity and viability measured on a BD Fortessa instrument (BD); more than 90% cells were viable and 95% of the cells were confirmed to express CD45. For each sample, one million cells were fixed for 22 hours at 4°C, quenched and stored at -80°C according to 10X genomic Fixation of Cells & Nuclei for Chromium Fixed RNA profiling (CG000478) using the Chromium Next GEM Single Cell Fixed RNA Sample preparation kit (PN-1000414, 10X Genomics, Pleasanton, CA). 250,000 cells per sample were used for probe hybridization using the Chromium Fixed RNA Kit, Mouse Transcriptome, 4rxn x 4BC (PN-1000496, 10X Genomics), pooled at equal numbers and washed following the Pooled Wash Workflow following the Chromium Fixed RNA Profiling Reagent kit protocol (CG000527, 10X Genomics). GEMs were generated on Chromium X (10X Genomics) with a target of 10,000 cells recovered and libraries prepared according to the manufacturer instructions (CG000527, 10X Genomics).