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Rad6 recruitment to the GAL1 promoter requires the Gal4 activator and the Bre1 E3 ligase. After a shift to YP + galactose medium, ChIP and conventional PCR analysis were carried out in a RAD6-HA HTB1 strain that contained wild-type genes and (A) gal4Δ (YCH001) or (B) bre1Δ (YCH002) deletions. The PCR reactions were electrophoresed on a 10% acrylamide gel, which was stained with ethidium bromide. All reactions were performed in the linear range of amplification.
