title: Analysis of internucleosomal DNA fragmentation in apoptotic thymocytes by dynamic sieving capillary electrophoresis.
abstract: The analysis, by slab gel electrophoresis, of internucleosomal DNA cleavage or laddering, characteristic of apoptosis in many cell systems, is labour intensive, difficult to automate and at best only semi-quantitative. In this report we show that CE, using dilute solutions of hydroxyethylcellulose as a replaceable sieving matrix, can be applied to the relatively rapid analysis of DNA laddering in whole digests of apoptotic rat thymocytes. Also, using the sensitivity of laser-induced fluorescence detection and the highly sensitive nuclei acid stain YO-PRO-1, the CE method reported here can use 1000-2000 fold fewer cells than needed for traditional slab gel methods.
authors.affiliation: Division of Chemical Pathology, University of Leicester, UK.
authors.forename: M D
authors.initials: MD
authors.lastname: Evans
authors.forename: J T
authors.initials: JT
authors.lastname: Wolfe
authors.forename: D
authors.initials: D
authors.lastname: Perrett
authors.forename: J
authors.initials: J
authors.lastname: Lunec
authors.forename: K E
authors.initials: KE
authors.lastname: Herbert
mesh_terms: D000818:Animals; D017209:Apoptosis; D002197:Capillary Action; D002458:Cell Fractionation; D004247:DNA / Q000032:analysis*; D004586:Electrophoresis / Q000379:methods*; D019286:Endopeptidase K; D008297:Male; D009707:Nucleosomes / Q000737:chemistry*; D051381:Rats; D011916:Rats, Inbred F344; D012260:Ribonucleases / Q000378:metabolism; D012697:Serine Endopeptidases / Q000378:metabolism; D013950:Thymus Gland / Q000737:chemistry* / Q000648:ultrastructure
chemical_list: D009707:Nucleosomes; D004247:DNA; D012260:Ribonucleases; D012697:Serine Endopeptidases; D019286:Endopeptidase K