title: GSM8122821: Input 1; Mus musculus; ChIP-Seq
library_construction_protocol: Primary hippocampal neurons, cultures were fixed on the plates and then scraped and resuspended in buffer B (50mM NaCl, 10mM Pipes pH6.8, 5mM MgCl 2 , 1mM CaCl 2 ), incubated for 5 min at 37_x000E_C and digested using 100U of MNase (Cell Signaling, 1011S) for 15 min at 37_x000E_C. MNase digestion was halted by adding EDTA to a final concentration of 5mM. SDS was added to a final concen- tration of 1% and samples were aliquoted and diluted with ChIP dilution buffer. Hippocampal samples were treated with 20ul of Protein G magnetic beads (Millipore, 16-662) and 1ul of H2A.Z (Millipore, ABE1348) antibody overnight at 4_x000E_C. The next day, samples were washed sequentially with low-salt (20mM Tris pH7.4, 0.1% SDS, 1% Triton X-100, 2mM EDTA, 150mM NaCl), high-salt (20mM Tris pH7.4, 0.1%SDS, 1% Triton X-100, 1mM EDTA, 500mM NaCl), LiCl (Millipore, 20-156) and TE (10mM Tris pH7.4, 1mM EDTA) buffers, and incubated while rotating for 5 min between washes. Immune complexes were extracted using TE Buffer and proteinase K (Roche, 3115828001) for both ChIP and Input samples, and heated at 65_x000E_C for 2 h, followed by 95_x000E_C for 10 min with QIAquick PCR Purification Kit (QIAGEN, 28104) for sequencing samples Purifed DNA was sent to Genome Quebec in Montreal, Canada where the library was constructed and sequenced on an Illumina HiSeq 4000