title: GSM7847723: whole tumour, scRNAseq; Mus musculus; RNA-Seq
library_construction_protocol: Mouse pancreatic tumours were dissected and placed into ice-cold DMEM/F-12 with 50 U/ml penicillin-streptomycin. Tissues were minced on ice and incubated in DMEM containing 1 mg/ml collagenase V  for 1 h at 37 °C. Digestion was stopped by adding ice cold G solution and cells were centrifuged (300 g, 5 min at 4 °C). Cells were further digested with TrypLE for 5 min at 37 °C, followed bystopping with ice-cold PBS with 2% FCS and centrifugation. Single cells were resuspended in PBS with 2% FCS and stained with fluorophore-conjugated antibody on ice in the dark when required. Cells were washed three times with PBS and filtered through a 40-μm nylon mesh and incubated in FACS buffer with 3 μM DAPI. Alive cells  were sorted with BD FACSymphony™ S6 (BD Biosciences) based on GFP or CD9 expression. Libraries were prepared using the Chromium Next GEM Single Cell 3' Reagent Kits v3.1 according to manufacturer's instructions.