Modification of comet-FISH technique by using temperature instead of chemical denaturation

Modification of comet-FISH technique by using temperature instead of chemical denaturation

Comet-FISH technique is an extension of commonly used comet assay. Its purpose is to determine whether primary DNA damage which comet assay detects occurred within a sequence of interest that is visualized by hybridization of fluorescent probe. Presence of the signal in comet tail indicates impaired...

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Bibliographic Details
Main Authors: Marin Mladinic, Davor Zeljezic
Format: Article
Language:English
Published: Elsevier 2014-01-01
Series:MethodsX
Subjects:
Comet-FISH
TP53
Temperature denaturation
Lymphocytes
Primary DNA damage
Gene fragmentation
Online Access:http://www.sciencedirect.com/science/article/pii/S2215016114200380
Description
Summary:Comet-FISH technique is an extension of commonly used comet assay. Its purpose is to determine whether primary DNA damage which comet assay detects occurred within a sequence of interest that is visualized by hybridization of fluorescent probe. Presence of the signal in comet tail indicates impaired structural integrity of sequence. Our modifications to the original comet-FISH technique described by Rapp et al. (2000) [1] include: • increase in probe binding specificity, • increased rate of successful hybridization, • simultaneous temperature denaturation of both, slide and probe.
ISSN:2215-0161

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