GSM6215859: Sample 1; Homo sapiens; RNA-Seq
Nuclei were isolated from frozen, dissected brain samples (50 - 60 mg) by dounce-homogenisation in nuclei extraction buffer. Homogenate was layered onto a sucrose cushion and ultra-centrifuged. Nuclei pellet was resuspended in resuspension buffer, filtered through a cell strainer cap, stained with DAPI and counted using a hemocytometer. Single nuclei RNA-seq libraries were prepared following the user guide of 10X Genomics (Chromium Single Cell 3' Reagents kit v3) with a target recovery of 10,000 nuclei per sample. Libraries were assessed using a High Sensitivity DNA Analysis Kit for the 2100 Bioanalyzer (Agilent), were then pooled equimolarily and treated with Illumina Free Adapter Blocking Reagent before sequencing on the NovaSeq 6000 System (Illumina).