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. 2021 Aug 17;3:673022. doi: 10.3389/fgeed.2021.673022

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Copyright © 2021 Atkins, Chung, Allen, Dampier, Gurrola, Sariyer, Nonnemacher and Wigdahl.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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DISCOVER-seq is a specialized version of ChIP-seq. As DSBs are introduced to DNA in living cells, endogenous repair processes recruit proteins to break sites. γH2AX localizes to DSBs within hundreds of base pairs in either direction. The MRN complex, recruited by γH2AX, localizes to the break site. Following genomic DNA extraction and fragmentation, antibody pull-down of fragments enables the sequencing of fragments surrounding DSBs. γH2AX pull-down is used in ChIP-seq and lacks the resolution to precisely locate DSB sites. MRN pull-down, specifically the MRE11 subunit of the MRN complex, enables precise sequencing of DSB sites with single-nucleotide resolution. Use of the MRE11 antibody for pull-down is the distinguishing characteristic of DISCOVER-seq compared to ChIP-seq. Created with Biorender.com.