title: Modification of comet-FISH technique by using temperature instead of chemical denaturation.
abstract: 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.
authors.affiliation: Institute for Medical Research and Occupational Health, Ksaverska cesta 2, 10000 Zagreb, Croatia.
authors.forename: Marin
authors.initials: M
authors.lastname: Mladinic
authors.affiliation: Institute for Medical Research and Occupational Health, Ksaverska cesta 2, 10000 Zagreb, Croatia.
authors.forename: Davor
authors.initials: D
authors.lastname: Zeljezic
keywords: Comet-FISH; Gene fragmentation; Lymphocytes; Primary DNA damage; TP53; Temperature denaturation