title: GSM8114726: Dclk1-ZSGreen+ cells from pancreas at 2 wks after KRAS activation, scRNAseq; Mus musculus; RNA-Seq
library_construction_protocol: Pancreas from Dclk1-DTR-ZSGreen, Mist1-Kras-TdTomato-Dclk1-DTR-ZSGreen mice at 2 weeks after KRAS activation and Mist1-Kras-TdTomato-Dclk1-DTR-ZSGreen mice16 weeks after KRAS activation were collected in ice-cold HBSS. Pancreas were dissociated with a scalpel and incubated in digestion medium (DMEM high glucose GlutaMAX, 1 mg/ml Collagenase P, 10 mM HEPES, Antibiotic-Antimycotic) at 37°C for 20 minutes with shacking. Isolated cells were then incubated in Accutase at 37°C for 5 minutes and filtered with 40 µm filters. Red blood cells were lysed by incubating cells in ACK lysing buffer on ice for 5 minutes. Cells were washed and resuspended in ice-cold PBS/0.5% BSA. DAPI was added for vital staining. Vital Dclk1 cells were sorted using FACS Aria II (Becton Dickinson) according to the presence of ZGreen and TdTomato signals using a C57BL/6-J mouse as negative control. Sorted cells were collected and immediately delivered for sequencing. Library was performed according to the manufacter's instructions (10x Genomics)