Break-seq
Double-Stranded Break Labeling
Break-seq combines DSB labeling with NGS to map chromosome breaks with improved sensitivity and resolution (Hoffman et al., 2015). DNA trapped in agarose plugs is end-repaired and labeled with biotin. The agarose is then digested by _-agarase, followed by dilution and fragmentation. The fragmented DNA is purified and end-repaired with unmodified dNTPs, followed by A-tailing. The fragments are purified on magnetic beads, PCR-amplified, purified, and used to prepare a sequencing library.
Advantages:
- Simple protocol
Disadvantages:
- Not replicated in other laboratories
Reagents:
Illumina Library prep and Array Kit Selector
Reviews:
None available yet
References:
Hoffman E. A., McCulley A., Haarer B., Arnak R. and Feng W. Break-seq reveals hydroxyurea-induced chromosome fragility as a result of unscheduled conflict between DNA replication and transcription. Genome Res. 2015;25:402-412