GSM7917407: pancreas, PanIN, scRNA-seq; Mus musculus; RNA-Seq
Mouse pancreases were digested into single-cell suspensions as previously described(Reichert, M. et al. Isolation, culture and genetic manipulation of mouse pancreatic ductal cells. Nat. Protoc. 8, 1354–1365 (2013).) Library was performed according to the manufater's instruction(10x Genomics).Briefly,GCs were resupended in the master mix 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 GEM was retrotranscripted to cDNA,which contains an illumina R1 primer sequence,Unique Molecular Identifier(UMI) and the 10x Barcode.The pooled barcoded cDNA was then cleaned up with Silane DynaBeads,anplified by PCRand the apropiated sized fragments were selected with SPRIselectreagent for subsequent library construction.During the library construction Ilumina R2 primer sequence,paired-end constructs with P5 and P7 sequence and a sampleindex were added.