title: The use of C0t-1 probe DNA for the detection of low levels of DNA fragmentation.
abstract: In many apoptotic systems the final demise of the DNA results in the generation of multinucleosomal-sized DNA fragments, which appear as a DNA ladder after agarose electrophoresis. Extensive DNA fragmentation can be detected by ethidium bromide staining. Visualization of low levels of DNA fragmentation, however, requires both a more sensitive detection system, as well as a method of DNA extraction, that limits the extent of high molecular weight DNA shearing. We have found that the use of C0t-1 repetitive DNA as a probe for Southern analysis of DNA ladders is a sensitive method to detect low levels of DNA fragmentation. We have applied this methodology in the detection of DNA fragmentation in the normal developing C57BL/6 mouse retina at stages in which there is known DNA fragmentation, as well as in human Y-79 retinoblastoma cells grown in culture. We have found that in many instances in which there is no detectable DNA ladder with ethidium bromide staining a very definitive DNA ladder can be visualized via Southern blot analysis with a C0t-1 DNA probe.
authors.affiliation: National Eye Institute, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: P
authors.initials: P
authors.lastname: Wong
authors.forename: S B
authors.initials: SB
authors.lastname: Smith
authors.forename: N
authors.initials: N
authors.lastname: Bora
authors.forename: S
authors.initials: S
authors.lastname: Gentleman
mesh_terms: D000818:Animals; D017209:Apoptosis; D002478:Cells, Cultured; D004247:DNA / Q000032:analysis*; D015342:DNA Probes; D018076:DNA, Complementary; D004273:DNA, Neoplasm / Q000032:analysis; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D014407:Tumor Cells, Cultured
chemical_list: D015342:DNA Probes; D018076:DNA, Complementary; D004273:DNA, Neoplasm; D004247:DNA