GSM7289740: Patient 2 (PDAC2), scRNAseq; Homo sapiens; RNA-Seq
Freshly resected human PDAC tissue was acquired in the grossing room immediately following resection via Whipple or distal pancreatectomy. Bulk tissue specimen was rapidly dissociated into single cell suspension to reduce transcriptional artifacts of handling and time. Subsequent lysis and single cell prep was performed as detailed previously Library was performed according to the manufacter's instructions (single cell 3' v2 protocol, 10x Genomics). Briefly, GCs were resuspended 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, which contains an Ilumina R1 primer sequence, Unique Molecular Identifier (UMI) and the 10x Barcode. 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. During the library construction Ilumina R2 primer sequence, paired-end constructs with P5 and P7 sequences and a sample index were added.