GSM8515672: PBMC CL307 +SubAB stimulated, rep 2; Homo sapiens; RNA-Seq
The scRNA-seq libraries were generated using Chromium Single Cell 30 Library & Gel Bead Kit v.3 (10x Genomics) according to the manufacturer's protocol. Briefly, cells were counted, diluted at 1,000 cells/mL in PBS and 20,000 cells were loaded in the 10x Chromium Controller to generate single-cell gel-beads in emulsion. After reverse transcription, gel-beads in emulsion were disrupted. Barcoded complementary-DNA was isolated and amplified by PCR. Following fragmentation, end repair, and A-tailing, sample indexes were added during index PCR. The purified libraries were sequenced on a Novaseq 6000 (Illumina) with 28 cycles of read 1, 8 cycles of i7 index and 91 cycles of read 2. scRNA-seq experiments were performed at the “Labtech SingleCell@Imagine” on PBMCs isolated from healthy blood donors' buffy coats. The scRNA-seq libraries were generated using Chromium Single Cell 30 Library & Gel Bead Kit v.3 (10x Genomics) according to the manufacturer's protocol. Briefly, cells were counted, diluted at 1,000 cells/mL in PBS and 20,000 cells were loaded in the 10x Chromium Controller to generate single-cell gel-beads in emulsion. After reverse transcription, gel-beads in emulsion were disrupted. Barcoded complementary-DNA was isolated and amplified by PCR. Following fragmentation, end repair, and A-tailing, sample indexes were added during index PCR. The purified libraries were sequenced on a Novaseq 6000 (Illumina) with 28 cycles of read 1, 8 cycles of i7 index and 91 cycles of read 2.