Detection of telomere damage as a result of strand breaks in telomeric and subtelomeric DNA.
Telomeres are the tandem repetitive DNA sequences at both ends of a chromosome with a repeating unit of TTAGGG. The integrity of a telomere is crucial to chromosomal stability and cellular viability. Damages to telomere DNA disrupt telomere integrity and accelerate telomere shortening. We describe a method for the assessment of strand breaks in the telomere/subtelomere region in cultured cells. Cells were embedded in agarose plugs and subjected to lysis and alkaline treatment to relax the DNA double helix. The telomere fragments as the result of strand breaks in the telomere/subtelomere region were then separated from the genomic DNA by electrophoresis, blotted onto membranes, and detected by a probe specific to the telomere sequence. Because of the large content of the telomere in human cells and the fact that telomere DNA is much more prone to damage than the bulk genomic DNA, the analysis may serve as a good indication of general DNA damage as well.
Li
Wei-guo
WG
Institute of Zoology, Chinese Academy of Sciences, Beijing, P. R. China. tanclswu@public.wh.hb.cn
Li
Qing-huan
QH
Tan
Zheng
Z
D004249:DNA Damage; D053938:DNA Fragmentation; D004260:DNA Repair; D004587:Electrophoresis, Agar Gel / Q000379:methods*; D006367:HeLa Cells; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D009838:Oligodeoxyribonucleotides / Q000302:isolation & purification; D016615:Telomere / Q000187:drug effects*
D009838:Oligodeoxyribonucleotides; D006861:Hydrogen Peroxide