title: GSM7783205: LLC, CT, scRNAseq; Mus musculus; RNA-Seq
library_construction_protocol: Tumors were gently minced into small pieces using scissors, and subsequently transferred into a 15 ml tube containing the digestion solution (2 mg/ ml Collagenase type IV (C5138, Sigma), 2.5 mg/ ml Dispase II (4942078001, Sigma), 40 µg/ ml DNase (10104159001, Sigma) in DMEM). Tumors were digested for 30 minutes at 37°C with continuous rotation. Following digestion, the resulting cell suspensions were filtered through 70 µm cell strainers (CSS010070, Jetbiofil) placed on 50 ml tubes. The cell suspensions were subsequently centrifuged at 300×g for 7 minutes at 4°C. The supernatants were aspirated completely. The cell pellets were re-suspended in 5 ml 1x ACK buffer on ice for 3 minutes to lyse the erythrocytes. Next, 8 ml PBS was added into the tubes to stop reaction. In the end, the cells were centrifuge at 300xg for 7 minutes at 4°C and resuspended with PBS for further analysis. Library was performed accoring to manufacter's instructions (single cell 3' v3.1 protocol, 10X Genomics)