GSM9349012: mPFC WT M Sample 525 ATAC; Mus musculus; ATAC-seq
Brain regions were dissected according to the Allen Brain Reference. Nuclei were isolated following the Miltenyi Biotec “Reliable nuclei extraction and enrichment” protocol. Dissected tissues were transferred to gentleMACS™ C Tubes with 2 mL ice-cold lysis buffer and dissociated using program 4C_nuclei_1. The suspension was filtered through a 70-μm strainer, rinsed with 2 mL lysis buffer, and centrifuged (300 × g, 5 min, 4 °C). Pellets were resuspended in resuspension buffer, passed through a 30-μm strainer, and centrifuged again (300 × g, 5 min). Nuclei were resuspended in 450 μL nuclei separation buffer per 10⁶ nuclei, mixed with 50 μL Anti-Nucleus MicroBeads, and incubated for 15 min at 4 °C. After adding 2 mL separation buffer, samples were subjected to magnetic separation. MS columns were placed on a MACS® Separator and equilibrated with 3 mL separation buffer. Labeled nuclei were loaded onto the column and washed twice with 1 mL separation buffer. Columns were removed from the magnet, placed in collection tubes, and the nuclei were eluted by adding separation buffer and pushing the plunger. 10x Chromium Single Cell Multiome ATAC + GEX library construction protocol recommended by manufacturer