학술논문

Protocol for mapping double-stranded DNA break sites across the genome with translocation capture sequencing
Document Type
article
Source
STAR Protocols. 4(2)
Subject
Biological Sciences
Bioinformatics and Computational Biology
Genetics
Human Genome
Biotechnology
1.1 Normal biological development and functioning
Underpinning research
Generic health relevance
Cell Biology
Genomics
High-throughput Screening
Language
Abstract
Translocation sequencing can be used to assess mechanisms of DNA repair and identify genome-wide double-strand breaks (DSBs) accessible to DNA repair machinery. Here, we present a protocol for mapping double-strand DNA break sites across the genome with translocation capture sequencing. Bait DSBs are introduced using a Cas9 nuclease and repaired by the host cell, connecting bait DSBs to other DSBs. Repair sites are detected by isolating bait site DNA, cleaving normal sequence to enrich off-site repair, and next-generation sequencing. For complete details on the use and execution of this protocol, please refer to Switonski et al. (2021).1.