Here we made use of a human osteosarcoma U-2 OS modified cell line (U-2 OS DIvA- Flag-GFP-SMC1), expressing a hydroxytamoxifen controlled restriction enzyme (AsiSI-ER which allows the induction of multiple annotated DSBs) and Flag GFP-SMC. We performed tandem purification before and after DSB induction followed by mass spectrometry to identify cohesin interacting partners.