GSM9082346: In vitro PDAC CAF cell line, untreated, donor 3; Homo sapiens; RNA-Seq
0.05% Accutase (trypsin) was used to harvest the cells. Libraries were constructed using the 10X Chromium single-Cell 3' reagent v3.1 and 5' reagent v2 kit per the manufacturer's protocol. Briefly, single-cell suspensions was mixed with 10X RT reaction master mix, and then loaded together with gel beads and partitioning oil into a 3' or 5' Chip. The chip was then loaded onto a chromium controller for single-cell GEM generation and barcoding. RNA transcripts were captured by poly(dT) primers and barcoded with cell barcodes and unique molecular identifiers. After emulsion disruption, cDNA molecules were amplified, followed by fragmentation, adapter, and sample indices addition. The final libraries were quantified with qubit assay, examined by Agilent Tape station, and sequenced by Illumina Novaseq 6000.