title: GSM8979111: AOM_DSS_Tumors_Mouse; Mus musculus; RNA-Seq
library_construction_protocol: AOM/DSS tumors were dissected and washed with HBSS (Gibco) supplemented with antibiotic antimycotic solution (Gibco). The tumors were minced and digested in in 1000 U/mL Collagenase IV (Sigma-Aldrich, St. Louis, MO, USA), 1 mg/mL Dispase II (Roche, Basel, Switzerland), and 100 U/mL Dnase I (Sigma-Aldrich, St. Louis, MO, USA) in DMEM (BioSera, France) in three serial 20-min digestions. Cell suspensions were centrifuged and washed in FACS buffer (5% FBS (Biowest) in PBS). 1–2 million cells/100 μL were incubated with anti-CD45 and anti-CD326 antibodies for the exclusion of immune and remaining epithelial cells, respectively, and 1 μg/mL Propidium Iodide (PI) (Sigma) for dead cell exclusion. Cell sorting was performed using the FACSAria III cell sorter (BD) and the FACSDiva (BD) software. For each sample, approximately 16,000 cells were loaded onto a 10X Genomics Chromium Controller using the Chromium Next GEM Single Cell 3' Kit v3.1 according to the manufacturer's protocol. Library preparation was performed following the 10X Genomics workflow, with quality control performed using an Agilent Bioanalyzer High Sensitivity DNA chip.