Protocol for mapping physiological DSBs using in-suspension break labeling in situ and sequencing

Osama Hidmi*, Sara Oster, Diala Shatleh, Jonathan Monin, Rami I. Aqeilan*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

Physiological double-stranded breaks (DSBs) are a major source of genomic instability. Here, we present a protocol for mapping physiological DSBs by in-suspension break labeling in situ and sequencing (sBLISS) in a single-nucleotide resolution. We describe steps for cell fixation, labeling of DSBs, DNA isolation followed by in vitro transcription (IVT), reverse transcription, and library preparation. sBLISS provides a map of DSBs over the genome and can be used to study the role of different factors in DSB formation. For complete details on the use and execution of this protocol, please refer to Hidmi et al.1

Original languageAmerican English
Article number103059
JournalSTAR Protocols
Volume5
Issue number2
DOIs
StatePublished - 21 Jun 2024

Bibliographical note

Publisher Copyright:
© 2024 The Authors

Keywords

  • Bioinformatics
  • Cancer
  • Cell Biology
  • Cell culture
  • Genomics
  • Molecular Biology
  • Sequence analysis

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