title: GSM3704199: anti-PD-1 treated tumor experiment 2; Mus musculus; RNA-Seq
library_construction_protocol: Mice were humanely euthanized, and mouse lungs were perfused using sterile PBS through heart perfusion from the left ventricle. The whole lung was cut and minced into small pieces followed by digestion in collagenase D (Sigma) and DNase I (Sigma) in Hank's Balanced Salt Solution (HBSS) at 37°C for 30 minutes. After incubation, the digested tissue was subjected to a 70-μm cell strainer (ThermoFisher Scientific) to obtain single-cell suspensions. Live cell swere then sorted using DAPI  staining. Cells were then resuspended into single cells at a concentration of 1 x 106 per ml in 1X PBS with 0.4% BSA for 10x genomics processing. Single Cell 3' Reagent Kits v2: Chromium™ Single Cell 3' Library & Gel Bead Kit v2, PN-120237; Single Cell 3' Chip Kit v2 PN-120236 and i7 Multiplex Kit PN-120262" (10x Genomics) scRNA-seq