GSM8993906: Mouse,PyMT,Tumor,Native Control,rep2; Mus musculus; RNA-Seq
Tumor tissues were minced and then digested at 37 °C for 40 min using an enzyme cocktail containing 150 U/mL of Collagenase Ⅳ, 20 U/mL of Dispase and 0.33 U/mL of DNase I, followed by filtering through a 40 μm strainer to obtain the single cell suspension. Acridine orange (AO) and propidium iodide (PI) were used for fluorescence staining of viable cells. Cell sorting was performed using the WOLF® Cell Sorter (NanoCellect). 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.