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Chromatin was extracted by the addition of 1 ml of NUN2 buffer containing 20 mM HEPES (pH = 7.6), 7.5 mM MgCl 2 , 0.2 mM EGTA, 300 mM NaCl, 1 M urea and 1 % NP-40, 1x protease and 1x phosphatase inhibitors and rotated at 4 C for 10 min

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The researchers used a central composite design with response surface methodology to find the best parameters for removal

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