GSM8831733: PDAC, TMAB3/siRNA, 7 days post-3rd treatment, rep2; Mus musculus; RNA-Seq
Pancreatic tumors were collected after 7 days post-treatment. To dissociate, tumors were minced and washed in Hank's Balanced Salt Solution (Corning) and incubated with collagenase (Collagenase type V 0.012% (w/v) (Sigma), Soy Trypsin Inhibitor 0.012% (w/v) (Gibco) and DNase (Sigma) and incubated with heating for 40 minutes at 37° C on the gentleMACS Octo Dissociator (Miltenyi Biotechnology) program (37_m_TDK_1). Cells were filtered using a 70 μm nylon mesh (Fisher Scientific). Ebioscience 1X RBC Lysis Buffer (Thermo Fisher) was added to lyse red blood cells. Single-cell suspensions were prepared, and dead cells were excluded using the EasyStep Dead Cell Removal (Annexin V) Kit (STEM Cell Technologies). For each group, 5,000 cells were used for the 10x Genomics 3' v2 single-cell assay. Reverse transcription, cDNA amplification, and library preparation were performed according to the manufacturer's protocol at the Yale Center for Genome Analysis.