GSM5419461: CM2260; Mus musculus; RNA-Seq
Microdissected lung tumors were dissociated with collagenase IV, dispase, and trypsin at 37 °C for 30 min. After dissociation, the samples were maintained on ice, in the presence of 2 mM EDTA and 1 U/mL DNase to prevent aggregation. For the sorting of viable cells, DAPI was used to exclude dead cells. FACSAria sorters (BD Biosciences) were used for cell sorting. Cells were loaded in each channel of Chromium Chip B (10X Genomics: 1000074) with a recovery target of 8,000 cells per sample, and emulsions were generated on the Chromium Controller (10X Genomics) Libraries were constructed using the Chromium Single Cell 3′ Library & Gel Bead Kit v3 kit (10X Genomics: 1000075).