Mapping of genome-wide DSBs in HEK293T cells using the RAFT procedure.

Large DNA fragments (mainly of 50-250 kb, denoted as forum domains) were isolated inside agarose plugs as described below in the extract protocol section. After ligation of 5'-bio-oligos to the blunt DNA termini the digestion with Sau3A enzyme was performed followed by selection of genome-wide preparation of forum domains termini on streptavidin-paramegnetic particles. After ligation of Sau3A adapter the termini were PCR amplified and used for deep sequencing. Overall design: Mapping of Double-Strand DNA Breaks/Forum Termini