Cohesin loading assays were done as described in (14 (link)) using the pUC19 plasmid. Topologically bound DNA-cohesin complexes were immunoprecipitated using a μMACS HA Isolation kit (Miltenyi Biotec). Following incubation with Pst I and/or protein digestion, the recovered DNA was analyzed by electrophoresis on a 0.8% (w/v) TAE agarose gel in 1× TAE and visualized as described above.
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