title: GSM5106120: AD1_SOX9pos; Homo sapiens; RNA-Seq
library_construction_protocol: Processing of tissue was completed on ice or at 4°C for the entirety of the protocol. In brief, ~100 mg of postmortem frozen human prefrontal cortex per donor was homogenized in 2 mL homogenization buffer (10 mM Tris pH 8, 250 mM sucrose, 25 mM KCl, 5 mM MgCl2, 0.1% Triton X100, 1% RNasin Plus, 1X Protease inhibitor, and 0.1 mM DTT in DNase- and RNase-free water) using a Wheaton Dounce tissue grinder (10 strokes with loose pestle, followed by 10 strokes with the tight pestle). Then, the homogenized tissue was filtered through 30 mm pre-separation filters to remove major debris. Samples were then centrifuged at 900 xg for 10 minutes at 4°C. Nuclei were then immunolabeled for SOX9. SOX9-positive or SOX9-negative nuclei were collected using a MoFlo XDP sorter with a 100 mm nozzle at 4°C. Collected nuclei were processed using the Single Cell 3' Gene Expression kit v3 (10x Chromium) according to manufacturer's instructions, and libraries were sequenced an Illumina NovaSeq 6000. Sequencing libraries were prepared using the Single Cell 3' Gene Expression kit v3 (10x Chromium) according to manufacturer's instructions.