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Micrococcal nuclease (MNase) digestion

SO Sungryong Oh
KB Kyungjin Boo
JK Jaebeom Kim
SB Seon Ah Baek
YJ Yoon Jeon
JY Junghyun You
HL Ho Lee
HC Hee-Jung Choi
DP Daechan Park
JL Ji Min Lee
SB Sung Hee Baek
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For making nucleosomes without H2BK12Ac or H2BK120ub, H2B-Flag WT/K12R/K120R were transfected into cells. Then, cells were lysed by MNase lysis buffer (50 mM Tris–HCl [pH 7.5], 1 mM CaCl2, 0.2% NP-40, Inhibitor Complex) and treated 50 U MNase (#M0247S, NEB) for making mono-nucleosomes. After MNase digestion, H2B-Flag containing mono-nucleosomes were pull down by Flag-M2 bead overnight, then these mono-nucleosomes were eluted by 3× Flag peptides (#F4799, Sigma).

Copyright and License information:  ©2020 The Author(s) 2020. Published by Oxford University Press on behalf of Nucleic Acids Research.
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