title: GSM8776220: WT1Cre TdTomDTR Depletion Orthotopic; Mus musculus; RNA-Seq
library_construction_protocol: Pancreas tissues/tumors were mechanically minced and enzymatically digested with collagenase V (1 mg/mL in Roswell Park Memorial Institute (RPMI) 1640) at 37°C. Normal Pancreas was incubated in Collagenase V for 15 minutes and orthotopic tumors for 25 minutes. Cell suspensions were filtered through 500 μm, 100 μm, and 40 μm mesh cell strainers. Dead cells were removed using the MACS Dead Cell Removal Kit (Miltenyi Biotec, catalog number 130-090-101). Single-cell complementary DNA libraries were prepared and sequenced at the University of Michigan Advanced Genomics Core using the 10× Genomics Platform Samples were run using 50-cycle, paired-end reads on the HiSeq 4000 (Illumina, San Diego, CA) to a depth of 100,000 reads.