Simultaneous Labeling of Single and DoubleStrand

Authors: José Luis Fernández Dioleyda Cajigal Jaime Gosálvez

Publish Date: 2011

Volume: , Issue: , Pages: 133-147

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Abstract

DNA Breakage DetectionFluorescence In Situ Hybridization DBDFISH permits simultaneous and selective labeling of single and doublestrand DNA breaks in individual cells either in the whole genome or within specific DNA sequences In this technique cells are embedded into agarose microgels lysed and subjected to electrophoresis under nondenaturing conditions Subsequently the produced “comets” are exposed to a controlled denaturation step which transforms DNA breaks into singlestranded DNA regions detected by hybridization with whole genome fluorescent probes or the probes to specific DNA sequences This makes possible a targeted analysis of various chromatin areas for the presence of DNA breaks The migration length of the DBDFISH signal is proportional to the number of double strand breaks whereas its fluorescence intensity depends on numbers of singlestrand breaks

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