title: GSM9001994: Mouse,PyMT,Lung,Chronic Stress,rep2; Mus musculus; RNA-Seq
library_name: GSM9001994
library_construction_protocol: The lung tissues were minced, and then digested at 37 °C for 30 min using an enzyme cocktail including 150 U/mL of Collagenase Ⅳ, 20 U/mL of Dispase, 5 U/mL of Elastase and 0.33 U/mL of DNase I. The cell solution was filtered through a 40 μm strainer to obtain single-cell suspension. Acridine orange (AO) and propidium iodide (PI) were used to stain living cells. WOLF® Cell Sorter (NanoCellect) was used for cell sorting. Sorted single cells were diluted in DMEM/F12 with 10% FBS, and loaded on a Chromium Controller to generate single-cell gel bead emulsions, targeting 20,000-30,000 cells for each tumor tissue (n=2 in the current study). Single-cell 3′ RNA-seq libraries were generated according to the manufacturer's instructions (Chromium Single Cell 3′ Reagent v2 Chemistry Kit, 10X Genomics, Inc.), followed by a sequencing analysis to reach an average depth of ~ 200,000 reads per cell using Illumina Novaseq 6000 system.
identifiers.namespace: GEO
identifiers.id: GSM9001994_r1