Breaks labeling and Enrichment on Streptavidin and Sequencing

BLESS is a genome-wide approach to map DSBs at nucleotide resolution (Crosetto et al., 2013). BLESS is able to detect telomere ends, Sce endonuclease_induced DSBs, and complex genome-wide DSB landscapes.In this method, DSBs are ligated in situ to a proximal linker covalently linked to biotin. The gDNA is extracted and fragmented, and the labeled fragments are captured on streptavidin beads. Next, a distal linker is ligated to the free end of the captured fragments. The fragments are released by linker digestion with I-SceI and PCR-amplified


  • Detects DSBs at nucleotide resolution
  • Does not depend on proteins that bind to DSBsãa source of bias
  • Does not depend on single-stranded DNA (ssDNA)ãa source of bias



Illumina Library prep and Array Kit Selector


Lee C. M., Cradick T. J., Fine E. J. and Bao G. Nuclease Target Site Selection for Maximizing On-target Activity and Minimizing Off-target Effects in Genome Editing. Mol Ther. 2016;24:475-487


Ran F. A., Cong L., Yan W. X., et al. in vivo genome editing using Staphylococcus aureus Cas9. Nature. 2015;520:186-191