PMID- 37549036
OWN - NLM
STAT- MEDLINE
DCOM- 20240308
LR  - 20240308
IS  - 2666-1667 (Electronic)
IS  - 2666-1667 (Linking)
VI  - 4
IP  - 3
DP  - 2023 Sep 15
TI  - exo-FISH: Protocol for detecting DNA breaks in repetitive regions of mammalian 
      genomes.
PG  - 102487
LID - S2666-1667(23)00454-9 [pii]
LID - 10.1016/j.xpro.2023.102487 [doi]
LID - 102487
AB  - Detecting DNA breaks in defined regions of the genome is critical to advancing 
      our understanding of genome stability maintenance. Here, we present exo-FISH, a 
      protocol to label exposed single-stranded DNA in defined repetitive regions of 
      mammalian genomes by combining in vitro restriction enzyme digestion on fixed 
      cells with fluorescence in situ hybridization (FISH). We describe steps for cell 
      harvesting and fixation, slide treatments, and FISH probe hybridization. We then 
      detail procedures for imaging and analysis. For complete details on the use and 
      execution of this protocol, please refer to Saayman et al. (2023).(1).
CI  - Copyright (c) 2023 The Authors. Published by Elsevier Inc. All rights reserved.
FAU - Saayman, Xanita
AU  - Saayman X
AD  - Sir William Dunn School of Pathology, University of Oxford, Oxford, UK. 
      Electronic address: xanitasaayman@gmail.com.
FAU - Graham, Emily
AU  - Graham E
AD  - Sir William Dunn School of Pathology, University of Oxford, Oxford, UK.
FAU - Leung, Chin Wei Brian
AU  - Leung CWB
AD  - Sir William Dunn School of Pathology, University of Oxford, Oxford, UK.
FAU - Esashi, Fumiko
AU  - Esashi F
AD  - Sir William Dunn School of Pathology, University of Oxford, Oxford, UK. 
      Electronic address: fumiko.esashi@path.ox.ac.uk.
LA  - eng
PT  - Journal Article
DEP - 20230806
PL  - United States
TA  - STAR Protoc
JT  - STAR protocols
JID - 101769501
RN  - 9007-49-2 (DNA)
RN  - 0 (DNA, Single-Stranded)
SB  - IM
MH  - Animals
MH  - In Situ Hybridization, Fluorescence/methods
MH  - *DNA/genetics
MH  - *Repetitive Sequences, Nucleic Acid/genetics
MH  - DNA, Single-Stranded
MH  - DNA Breaks
MH  - Mammals/genetics
PMC - PMC10425934
OTO - NOTNLM
OT  - Cell Biology
OT  - In Situ Hybridization
OT  - Microscopy
OT  - Molecular Biology
OT  - Single Cell
COIS- Declaration of interests The authors declare no competing interests.
EDAT- 2023/08/07 19:10
MHDA- 2024/03/08 06:43
PMCR- 2023/08/06
CRDT- 2023/08/07 12:35
PHST- 2023/04/14 00:00 [received]
PHST- 2023/06/07 00:00 [revised]
PHST- 2023/07/12 00:00 [accepted]
PHST- 2024/03/08 06:43 [medline]
PHST- 2023/08/07 19:10 [pubmed]
PHST- 2023/08/07 12:35 [entrez]
PHST- 2023/08/06 00:00 [pmc-release]
AID - S2666-1667(23)00454-9 [pii]
AID - 102487 [pii]
AID - 10.1016/j.xpro.2023.102487 [doi]
PST - ppublish
SO  - STAR Protoc. 2023 Sep 15;4(3):102487. doi: 10.1016/j.xpro.2023.102487. Epub 2023 
      Aug 6.