title: GSM4776917: ATAC-seq_female_E2_2; Mus musculus; ATAC-seq
library_construction_protocol: Tissue (4 animals pooled per replicate) was homogenized 15x with a loose pestle in a glass homogenizer containing Homogenization Medium (250 mM sucrose, 25 mM KCl, 5 mM MgCl2, 20 mM Tricine-KOH, 1 mM DTT, 0.15 mM spermine, 0.5 mM spermidine, 1X Roche EDTA-free protease inhibitor cocktail, pH 7.8). 0.3% IGEPAL CA-630 was added, and the tissue was further dounced 5x with a tight pestle. After douncing, the homogenate was filtered through a 40 µm strainer and then centrifuged at 500 x g for 15 min at 4degC. The pellet was resuspended in 0.5 ml homogenization buffer containing 1 mM DTT, 0.15 mM spermine, 0.5 mM spermidine, and 1X EDTA-free PIC. 30,000 GFP+ nuclei were collected into cold ATAC-RSB (10 mM Tris-HCl pH 7.5, 10 mM NaCl, 3 mM MgCl2) using the Sony SH800S Cell Sorter (purity mode) with a 100-?m sorting chip. After sorting, 0.1% Tween-20 was added, and the nuclei were centrifuged at 500 x g for 5 min at 4degC. The nuclei pellet was directly resuspended in transposition reaction mix. Nextera DNA Library Prep Kit