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.

Keywords

• Comet-FISH
• TP53
• Temperature denaturation
• Lymphocytes
• Primary DNA damage
• Gene fragmentation