GSM7433179: Donor1_DissociatedCNScells; Homo sapiens; RNA-Seq
High quality brain cells were used to generate single cell RNA-seq libraries using the Chromium Next GEM Single Cell 3' reagent dual index kit v3.1 following manufactures protocol (CG000315 • Rev E). Diluted cell suspensions (>90% viability) with a targeted recovery of 10000 cells were barcoded using GEM technology (Gel Beads-in-emulsion) in a chromium controller device. Library preparation performed according to manufacturer's protocol. First-strand synthesis was performed within GEMs before droplet breaking and clean-up. cDNA was amplified for 11 cycles and cleaned up. Amplified cDNA was then fragmented, end-repaired, A-tailed, and ligated to Illumina sequencing adapters. Adapter PCR was performed with a dual-index strategy. Dual-indexed libraries were sequenced by Illumina Nextseq 2000 with the following conditions: Read 1 - 28 cycles; Read 2 - 90 cycles; i7 - 10 cycles; i5 – 10 cycles.