GSM8147269: S4, Hippocampus, wild-type, 12m; Mus musculus; RNA-Seq
We adapted our single nuclei isolation for mouse hippocampus samples from the previously described protocol for rats in Phillips et al. Cell Reports 2022 and sorted nuclei at the University of Alabama at Birmingham's (UAB) Comprehensive Flow Cytometry Core (CFCC) using fluorescence-activated cell sorting (FACS) on an Atlas BD ARIA II. Per the standard 10X protocol, the UAB CFCC used the 10X Chromium Controller to partition hippocampal nuclei into droplets with a barcoded gel bead where nuclei were lysed, and RNA was reverse transcribed, creating cDNA encapsulated in each droplet. After fragmenting and amplifying cDNA, they added Illumina adapters using the Chromium Single Cell 3ʹ GEM, Library & Gel Bead Kit v3 (10X#: PN-1000121). The UAB Heflin Center for Genomic Sciences at the UAB Sequencing Core indexed and sequenced all samples on a NovaSeq 6000 (Illumina). For each sample, they loaded approximately 20,000 nuclei onto an S4 flow cell.