GSM8354307: TCH4-TCH6, D and R (SC); Homo sapiens; RNA-Seq
Single-cell samples were profiled by  10x chromium Single Cell RNA sequensing. Single-cell gene expression libraries were prepared according to the Chromium NextGEM Single Cell Gene Expression 3v3.1 kit (10x Genomics). Briefly, single cells, reverse transcription reagents, gel beads containing barcoded oligonucleotides, and oil were loaded on a Chromium controller (10x Genomics) to generate single-cell GEMs (Gel Bead-In-Emulsion). Full-length cDNA was synthesized and barcoded within each GEM. Subsequently, GEMs were broken, and cDNA was pooled. Following cleanup using Dynabeads MyOne Silane Beads, cDNA was amplified by PCR. The amplified product was fragmented prior to end-repair, A-tailing, and adaptor ligation. Patient samples were flow-sorted, gating for CD45-dim and SSC low cells; 10K–15K cells were then labeled using the 10x Genomics (Pleasanton, CA) Chromium Next GEM Single Cell 3' Kit v3.1 and sequenced at 200 M reads per sample using an Illumina NovaSeq 6000.