GSM8252443: Vec1; Mus musculus; RNA-Seq
freshly collected pancreatic orthotopic tumors from C57BL/6 mice injected with KPC-OE-Vector cell or KPC-OE-MLKL cells were flushed with ice-cold PBS  then performed by  digestor. The RBC cells were removed and the isolated cells were resuspended in digestion buffer (HBSS with Mg2+/Ca2+ supplemented with 2mg/ml collagenase type I and 40 U/ml DNase I) and incubated at 37℃ for 30min. Cells were washed and re-suspended in PBS and stained with a vialility dye to evalutaed the cell viability. library was performed according to the manufacter's instructions (single cell 3'v2 protocol, 10x Genomics). Briefly, cells were respended in the master mix and loaded together with partitioning oil and gel beads into the chip to generate the gel bead-in-emulsion(GEM). the poly-A RNA from the cell lysate contained in every single GEM was retrotranscripted to cDNA. the pooled barcoded cDNA was then cleaned up with silane dynabeads, amplified by PCR and the apropiated sized fragments were selected with SPRIselect reagent for subsequent library construction.