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Subject: chromatin shearing biol. Chromatin Immnunoprecipitation (ChIP)
ChIP assays were performed according to the ChIP-IT Express instruction manual (Active Motif, catalog # 53008). Cells were plated at 15x106 cells per 15 cm culture dish in cMEM for 2 d, then serum starved in modified IMEM for 2 d. Cells were treated with R5020 (10–8 M) or vehicle for 1 or 4 h. For T47D cells expressing inducible PR, AP21967 (10–9M) was added during the starvation step. "Chromatin was sheared using a Bioruptor sonicator "(Diagenode, model UCB-200), for 30 min (30 s on/off).Спасибо!
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