GSM7818312: ATAC-Seq - Hippocampus Aged Male Replicate 3; Mus musculus; ATAC-seq
The hippocampus was homogenized with a Dounce tissue grinder (Wheaton 357544) in 2mL HB buffer (1 mM DTT, 0.15 mM spermine, 0.5 mM spermidine, protease inhibitor (Sigma-Aldrich 04693159001), 0.3% IGEPAL-630, 0.25 M sucrose, 25 mM MgCl2, 20 mM Tricine-KOH). The homogenate was then filtered through a 40 mm strainer. The filtrate was centrifuged over one volume of cushion buffer (0.5 mM MgCl2, 0.5 mM DTT, protease inhibitor, 0.88 M sucrose) at 2800 g for 10 minutes in a swinging bucket centrifuge at 4°C. The nuclei pellet was resuspended in 20 mL PBS. 1 mL of the nuclei sample was stained with Hoechst 33342 to calculate nuclei concentration, and 50,000 nuclei per sample were used for downstream library preparation. Libraries were prepared using the Nextera DNA Library Prep kit (Illumina FC-121-1030) and quality was analyzed using the D1000 ScreenTape Assay (Agilent 5067-5582).