title: Single-cell RNA-Sequencing of pancreatic and PDAC stromal cells
design: Single-cell RNA-Sequencing of pancreatic and PDAC stromal cells
library_construction_protocol: Digested xenograft, electroporated tumor and mice pancreas cells were stained with a total of 3 fluorescent cell surface antibodies and DAPI. Cells were sorted into four populations according to CD326+ (EPCAM+), EPCAM-/CD31+, EPCAM-/CD45+ and EPCAM-/CD31-/CD45- expression. Cells from each population were sorted in PBS+5%BSA and mixed in a 2:1:1:1 ratio. 30.000 cells of each sample were further processed for single cell analysis. The single cell analysis of full tumor and pancreas was carried out with 10X Genomics Chromium Single Cell Kit Version 3,. Suspensions were prepared as described above and diluted in PBS, 30.000 cells were used as input and added to 10x Chromium RT mix. For downstream cDNA synthesis (12-14 PCR cycles), library preparation, and sequencing, we followed the manufacturer's instructions.  The single cell analysis of full tumor and pancreas was carried out with 10X Genomics Chromium Single Cell Kit Version 3,. Suspensions were prepared as described above and diluted in PBS, 30.000 cells were used as input and added to 10x Chromium RT mix. For downstream cDNA synthesis (12-14 PCR cycles), library preparation, and sequencing, we followed the manufacturer's instructions.