GSM8795024: Pulmonary pleomorphic carcinoma, scRNA-seq, multiplexed; Homo sapiens; RNA-Seq
Fresh frozen PPC tissue samples (25 mg) from four patients were fixed and dissociated using the Chromium Next GEM Single Cell Fixed RNA Sample Preparation Kit (10x Genomics) according to the manufacturer's protocol. Fixed single-cell suspensions were hybridized with Probe Barcodes from the Chromium Fixed RNA Kit, Human Transcriptome (10x Genomics), for 19 hours at 42°C. After hybridization, single-cell suspensions were pooled with an equal number of cells from each sample. Gel Beads-in-Emulsion (GEMs) were generated using the Chromium Controller (10x Genomics). GEMs were processed for probe ligation, pre-amplified by PCR, and cleaned using SPRIselect beads (Beckman Coulter). Gene expression libraries were constructed using the Chromium Fixed RNA Profiling Reagent Kits for Multiplexed Samples (10x Genomics). Sample Index PCR was performed with the Dual Index Kit TS Set A (10x Genomics) to add Illumina adapters and indexes. Final libraries were purified, quality-checked and sequenced on the Illumina NovaSeq 6000 platform (150 bp paired-end).