GSM5526330: Patient 1 Prefrontal cortex sample 1; Homo sapiens; RNA-Seq
Nuclei were isolated from fresh frozen autopsy tissue following the 10x Genomics (Pleasanton, CA) recommended protocol using 0.1% Nonident NP40 lysis buffer. Suspensions were filtered through a 40 um mesh to remove debris and concentrated to a density of 1x10^6 nuclei/mL in Dulbecco's PBS + 0.04% BSA (Sigma Aldrich). Nuclei were processed according to Chromium 3' Gene Expression V3 Kit (10X Genomics, Pleasanton, CA) using manufacturer's guidelines. An estimated 15,000 nuclei were loaded onto each channel with a targeted cell recovery of 10,000 nuclei/sample. This was followed by sequencing on a S4 NovaSeq chip (Illumina Inc., San Diego, CA). Qubit 3 (Fisher Scientific) and 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA) were used for quality check of cDNA. The output BAM file, a binary text file containing the sequence alignment data from sequencing, was processed through 10X Genomics Cell Ranger software v3.1.0. Sequencing generated scRNA-seq data with ~41,000 2x100bp reads per nucleus and 63,653 total cells sequenced