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Cell lysates were centrifuged at 2300 x g for 5 min at 4 C to isolate the nuclei. Nuclei were suspended in 500 μL of 0.5% SDS lysis buffer 0.5% SDS 10 mM EDTA 50 mM Tris-HCl pH 8.0 and subjected for soni ion to shear chromatin fragments to an average size between 200bp and 500bp on the Branson Sonifier 450 ultrasonic processor 20% amplitude 0.5 s on 1 s off for 30 s .
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