An improved method for the detection of DNA fragmentation

A. Facchinetti, L. Tessarollo, M. Mazzocchi, R. Kingston, D. Collavo, G. Biasi

Research output: Contribution to journalArticle

Abstract

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.

Original languageEnglish
Pages (from-to)125-131
Number of pages7
JournalJournal of Immunological Methods
Volume136
Issue number1
DOIs
Publication statusPublished - Jan 24 1991

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Keywords

  • Apoptosis detection
  • DNA fragmentation
  • Southern blot analysis

ASJC Scopus subject areas

  • Biotechnology
  • Immunology

Cite this

Facchinetti, A., Tessarollo, L., Mazzocchi, M., Kingston, R., Collavo, D., & Biasi, G. (1991). An improved method for the detection of DNA fragmentation. Journal of Immunological Methods, 136(1), 125-131. https://doi.org/10.1016/0022-1759(91)90258-H