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