title: GSM9656713: Sample 5 single cell for microglia sequenced with novaseq; Mus musculus; RNA-Seq
library_name: GSM9656713
library_construction_protocol: For single nuclei data: Hippocampal tissue was dissociated in ice-cold HBSS and mechanically homogenized. Nuclei were filtered, washed, and purified through sequential centrifugation steps. The final nuclei suspension was resuspended in RNase inhibitor-containing buffer, filtered, and counted prior to loading for single-nucleus RNA-seq. For single cell data: Hippocampal tissue was dissociated in ice-cold HBSS and mechanically homogenized. Cell suspensions were filtered, subjected to debris removal, and microglia were enriched using CD11b magnetic bead-based isolation. Purified cells were resuspended in RNase inhibitor-containing buffer and assessed for viability prior to scRNA-seq. Single-nucleus and single-cell RNA-seq libraries were generated using the Chromium™ Single Cell 3′ Library & Gel Bead Kit v3.1 (10x Genomics, PN-1000121) following the manufacturer's instructions, targeting recovery of up to 10,000 nuclei per sample. For single nucleus data, twenty-four libraries were pooled and sequenced on one NextSeq 500 flow cell and four NovaSeq 6000 SP flow cells (Illumina) with 28 bases for read 1, 55 bases for read 2, and 8 bases for index read 1. For single cell data, twenty-four libraries from different animals were pooled based on molar concentrations and sequenced on one NextSeq 500 flow cell and two NovaSeq 6000 SP flow cells (Illumina) with 28 bases for read 1, 55 bases for read 2, and 8 bases for index read 1.
identifiers.namespace: GEO
identifiers.id: GSM9656713_r1