GSM5492904: PIM1 AR 1; Homo sapiens; ChIP-Seq
Cell pellets were resuspended in nuclei isolation buffer (50 mM Tris–pH 8.0, 60 mM KCl, 0.5% NP40), nuclei collected, and resuspended in sonication buffer (RIPA buffer). Samples were sonicated in TPX PMP tubes (Diagenode) for 60 min (30 sec. on, 30 sec. off) in a Bioruptor sonicator (Diagenode). Inputs (10%) were collected and supernatants were then incubated overnight with the corresponding antibodies pre-incubated with Protein A and Protein G Dynabeads (Invitrogen). Immunocomplexes were then washed and cross-linking reversed overnight at 65 °C with 5 M NaCl. DNA was isolated with the Zymo Chip DNA Clean and Concentrator kit (Zymo Research). Libraries were generated using NEBNext Ultra II DNA Library Prep Kit.