GSM6807291: Mixture5; Homo sapiens; RNA-Seq
Single-cell samples were submitted to the Baylor College of Medicine Single Cell Genomics Core immediately after preparation. Per sample, 16K cells were loaded, with an expected return of 10K cells. 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. Final libraries were generated by amplification. Sequencing of single-cell libraries was performed by the Genomics and RNA Profiling Core at Baylor College of Medicine. To reach an estimated 20K reads per cell, samples were sequenced at a depth of 200M reads on the NovaSeq 6000 platform (Illumina).