A rapid and efficient method for preparation of genomic DNA suitable for analysis of both high and low molecular weight DNA fragmentation during neuronal apoptosis.
In this paper, we describe a fast (within 1 h) and efficient method for the preparation of intact genomic DNA suitable for the analysis of DNA integrity in neuronal cells undergoing apoptosis. The procedure involves the embedding of apoptotic cells into low-melting point agarose, in situ cell lysis and purification of DNA samples followed by resolution of fragmented DNA by agarose gel electrophoresis. The method has no restrictions on the number of simultaneous DNA preparations, allows detection of both high and low molecular weight DNA fragments, and is suitable for the preparation of DNA samples from cultured cells, fresh or frozen tissues, as well as from purified nuclei. This procedure is also applicable for the preparation of intact genomic DNA for the megabase-scale analysis.
Solovyan
V
V
Department of Neuroscience and Neurology, University of Kuopio, P.O. Box 1627, FIN-70211, Kuopio, Finland.
Salminen
A
A
D000818:Animals; D017209:Apoptosis; D002474:Cell-Free System; D053938:DNA Fragmentation; D004273:DNA, Neoplasm / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification*; D016678:Genome; D051379:Mice; D008967:Molecular Biology / Q000379:methods; D008970:Molecular Weight; D009447:Neuroblastoma; D009474:Neurons / Q000166:cytology* / Q000502:physiology; D014407:Tumor Cells, Cultured
D004273:DNA, Neoplasm