GSM8441219: GFP_H3K27ac_LPS1; Mus musculus; ChIP-Seq
Only for the Nur77 ChIP, cells were crosslinked directly in culture flasks with 2mM Di(N-succinimidyl) glutarate in PBS for 30 min at room temperature. For all ChIP targets including Nur77, cells were then crosslinked with 1% formaldehyde for 10 minutes at room temperature. After quenching and cell lysis, the chromatin was sonicated into approximately 400bp fragments using a Bioruptor Pico (Diagenode) for 4 cycles (double cross-linked) or 3 cycles (single cross-linked) of 10sec on and 50sec off. Immunoprecipitation was performed using HA (for Nur77),  RNA polymerase, or H3K27ac antibody Strrand-specific ChIP seq libraries were prepared using the KAPA hyperprep kit protocol. Concentration and fragment size of chromatin was confirmed using the Tapestation4200 (Agilent).