GSM6532915: KPC46 vehicle control 1; Mus musculus; RNA-Seq
KPC tumors were isolated from 4 mice, two treated with TAK-981 and two control mice. Tumors were dissociated using the mouse Tumor Dissociation Kit (Miltenyi) and the gentleMACS Octo Dissociator (Miltenyi), followed by 70 µm cell strainer filtering with 10 ml of cell wash buffer (1x PBS + 0.04% BSA) and centrifugation at 300 relative centrifugal force (rcf) for 5 min at room temperature. The pellets were treated with 1 ml ACK lysing buffer (Gibco) for 1 min, diluted with 10 ml wash buffer, centrifuged at 300 rcf for 5 min at 4°C, and resuspended with 10-15 ml wash buffer (aiming for 1,500 cells/µl). Cells were counted using Thermo Fisher Countess 3 Cell Counter and 10,000-20,000 total live cells were prepared for each sample. For cells with less than 70% viability, dead cell removal was performed with aim for >80% viability for all samples using Dead Cell Removal Kit (Miltenyi). Library was performed according to the manufacter's instructions (Single Cell 3' v3.1 Chemistry Dual index, 10x Genomics).