An improved method for the detection of DNA fragmentation.
An application of the Southern blot technique is described which permits the detection of DNA fragmentation due to cell death by apoptosis. DNA fragments were isolated from cell suspensions and tissues, separated on agarose gel, transferred by Southern blot and hybridized with a radiolabeled total cellular DNA probe. The application of this procedure to thymus cell samples, revealed the distinct ladder pattern of DNA fragments in multiples of about 180-200 base pairs, a characteristic feature of DNA fragmentation. In comparison to conventional DNA visualization with ethidium bromide staining, the radiolabeled probe improved the detection of DNA fragments at least eight-fold. This method detects low levels of DNA fragments, as well as physiological tissue DNA fragmentation, while avoiding cell damage due to DNA radiolabeling.
Facchinetti
A
A
Chair of Immunology, Institute of Oncology, University of Padua, Italy.
Tessarollo
L
L
Mazzocchi
M
M
Kingston
R
R
Collavo
D
D
Biasi
G
G
D000818:Animals; D002470:Cell Survival; D004247:DNA / Q000378:metabolism*; D004996:Ethidium; D051379:Mice; D008810:Mice, Inbred C57BL; D009693:Nucleic Acid Hybridization; D013194:Staining and Labeling
D004247:DNA; D004996:Ethidium