title: GSM8616984: PYMT; Mus musculus; RNA-Seq
library_construction_protocol: Tumor tissues were manually minced and transferred to digestion buffer. Digestion buffer was prepared by adding collagenase D (Worthington Biochem) (400U/mL for PyMT tumors or 500U/mL for KPC tumors), 20 μg/mL DNase I (Sigma), and 2% fetal bovine serum (FBS), into Hanks Balanced Salt Solution (HBSS) (Thermo Fisher). Tumors were digested for 30 min at 37C with constant stirring, then quenched by ethylenediaminetetraacetic acid (EDTA), filtered through 40 μm Nylon mesh, pelleted by centrifugation (750 g for 5 min at 4C), and suspended in phosphate buffered saline (PBS) for staining. Endothelial cells were sorted and processed for scRNAseq using Chromium Single Cell 3' Library and Gel Bead Kit v2 (10x Genomics) according to manufacturer guidelines.