PMID- 23913296
OWN - NLM
STAT- MEDLINE
DCOM- 20140317
LR  - 20130805
IS  - 1940-6029 (Electronic)
IS  - 1064-3745 (Linking)
VI  - 1054
DP  - 2013
TI  - Fluorescence in situ hybridization on electrophoresed cells to detect sequence 
      specific DNA damage.
PG  - 219-35
LID - 10.1007/978-1-62703-565-1_15 [doi]
AB  - Fluorescence in situ hybridization (FISH) to label fragments of DNA with probes 
      which can specifically locate a genomic region of interest, combined with the 
      single cell electrophoresis (Comet) assay, also termed Comet-FISH, allows the 
      quantification of DNA damage and repair at a specific genomic locus. While the 
      Comet assay alone quantifies only the overall DNA damage of an individual cell, 
      subsequent FISH on the electrophoresed single cell genome enables the 
      coincidental localization of fluorescently labelled sequences (i.e., probes) to 
      the respective damaged or undamaged genes or specific genomic regions of 
      interest. In that way sequence specific DNA damage, global genomic and 
      transcription coupled repair or the three dimensional ultrastructure of cells 
      from any tissue can be comparatively investigated. This protocol provides a 
      detailed description of the principles and basic methodology of a standard 
      Comet-FISH experiment to study interphase cells of any tissue. Also important 
      variations of the protocol (e.g., neutral conditions to detect double strand 
      breaks) as well as the production of fluorochrome-labelled DNA probes via random 
      priming are described.
FAU - Laubenthal, Julian
AU  - Laubenthal J
AD  - School of Life Sciences, University of Bradford, Bradford, UK.
FAU - Anderson, Diana
AU  - Anderson D
LA  - eng
PT  - Journal Article
PL  - United States
TA  - Methods Mol Biol
JT  - Methods in molecular biology (Clifton, N.J.)
JID - 9214969
RN  - 0 (DNA Probes)
RN  - 9007-49-2 (DNA)
SB  - IM
MH  - Comet Assay/*methods
MH  - DNA/*chemistry
MH  - *DNA Damage
MH  - DNA Probes/chemistry
MH  - Humans
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Staining and Labeling
EDAT- 2013/08/06 06:00
MHDA- 2014/03/19 06:00
CRDT- 2013/08/06 06:00
PHST- 2013/08/06 06:00 [entrez]
PHST- 2013/08/06 06:00 [pubmed]
PHST- 2014/03/19 06:00 [medline]
AID - 10.1007/978-1-62703-565-1_15 [doi]
PST - ppublish
SO  - Methods Mol Biol. 2013;1054:219-35. doi: 10.1007/978-1-62703-565-1_15.