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Chromatin fragmentation

XH Xiaorong Huang
QP Qinwei Pan
YL Ying Lin
TG Tingting Gu
YL Yi Li
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Incubate the nuclei with 1 μl of micrococcal nuclease (MNase) for 10 min in a water bath set at 37 °C. Gently invert the tubes every 2 or 3 min during incubation (before this, appropriate digestion time needs to be determined for each chromatin preparation.)

Transfer the tubes into ice box and add 20 μl of 0.5 M EDTA to each tube to stop the reaction.

Centrifuge the tubes at 13,000 g for 10 min at 4 °C.

Transfer the supernatant (S1, about 250 μl each tube) into a new 5 ml Eppendorf tube on ice. Re-suspend the pellet in 250 μl of lysis buffer. Leave the tube on ice to dialyze for 1 h.

Centrifuge the samples at 13,000 g for 10 min at 4 °C. Transfer the supernatant (S2, about 250 μl each tube) and combine with S1 tube, mix them on ice (500 μl in total each tube).

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