Protocol for mapping physiological DSBs using in-suspension break labeling in situ and sequencing.
STAR Protoc
; 5(2): 103059, 2024 Jun 21.
Article
em En
| MEDLINE
| ID: mdl-38717906
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.
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Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Quebras de DNA de Cadeia Dupla
Limite:
Humans
Idioma:
En
Ano de publicação:
2024
Tipo de documento:
Article