Nucleosome reconstitution

MJ Muhammad Jbara
SL Shay Laps
MM Michael Morgan
GK Guy Kamnesky
GM Guy Mann
CW Cynthia Wolberger
AB Ashraf Brik
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Histone octamers were reconstituted and purified as described previously52 from recombinant Xenopus Laevis histone sequences (H2AK119DHAUb, H2B, H3, and H4). The octamers bearing the H2AK119DHAUb were then reconstituted into nucleosomes with salt gradient dialysis and purified using a DEAE-5PW HPLC column (TOSOH). After purification, nucleosomes were dialyzed into NCP storage buffer (10 mM Tris-HCl pH 7.5, 50 mM KCl) and stored at 4 °C.

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