GSM6739144: hTERT-HPNE, scRNAseq; Homo sapiens; RNA-Seq
Panc04.03 and hTERT-HPNE were detached from culture dish using TrypLE (ThermoFischer) followed by washing and counting of the cell. PBMC were thawed in water bath followed by washing with warm RPMI-1640 media containg 10% FBS and counting. Library was performed according to the manufacter's instructions (Chromium Next GEM Single Cell 5` Gel Bead Kit v2 protocol, 10x Genomics). DNA-barcoded lectin reacted cells 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 and DNA-barcodes were amplified, which contains an Ilumina R1 primer sequence, Unique Molecular Identifier (UMI) and the 10x Barcode. The pooled barcoded cDNA and DNA-barcode was then cleaned up with Silane DynaBeads, amplified by PCR and the apropiated sized fragments were selected with SPRIselect reagent for subsequent library construction using Dual Index Plate TN Set A and Dual Index Plate TT Set A for cDNA and DNA-barcode respectively.