title: GSM2794699: MNase_WT_2wk_Cortex; Mus musculus; MNase-Seq
library_construction_protocol: Tissue were homogenized in L1 buffer (50mM Hepes pH7.5, 140mM NaCl, 1mM EDTA, 1mM EGTA, 0.25% Triton X-100, 0.5% NP40, 10% Glycerol, protease inhibitors) and spun down. Pellets were resuspended in L2 buffer (10mM Tris-HCl pH8.0, 200mM NaCl) and rotated at room temperature for 10 min. Tubes were spun down and the pellet was resuspended in 100 µl MNase digestion buffer (20mM Tris-Cl, pH 7.5, 10mM NaCl, 2.5mM CaCl2, 0.01 mM PMSF, 1x Roche protease inhibitors) and incubated with 40µg of RNase A (Qiagen), and incubated at room temperature for 10 min. Treated samples with 1 µl MNase (NEB) for 4min at 37C. The reaction was stopped with addition of 2 µl 0.5M EDTA. Triton X-100 was added to a final concentration of 1%, and samples were rotated at 4°C for 30 min. DNA was subsequently purified using Qiagen PCR purification kit. Libraries were constructed by using Ovation Ultralow Library Systems (Nugen) following manufacturer instructions.