A new approach for the electrophoretic detection of apoptosis

Nucleic Acids Res. 1996 Oct 15;24(20):4092-3. doi: 10.1093/nar/24.20.4092.

Abstract

Apoptotic cell death is often characterized by internucleosomal cleavage of genomic DNA, which exhibits a distinctive ladder upon electrophoresis. However, techniques used for the isolation and detection of DNA to demonstrate laddering may not be sufficiently sensitive, particularly when cleaved DNA is present at modest levels. We propose a new approach for isolating total cellular DNA using a silica-based resin that improves the resolution of DNA laddering. In addition, we introduce a rapid DNA labeling method that can increase the sensitivity of detecting DNA laddering. Each of these methods can be used for DNA from cell cultures or tissues.

MeSH terms

  • Animals
  • Apoptosis*
  • Brain / metabolism
  • Cells, Cultured
  • Cerebellum / metabolism
  • DNA / analysis*
  • DNA / metabolism
  • DNA Fragmentation
  • DNA-Directed DNA Polymerase / metabolism
  • Electrophoresis, Agar Gel / methods*
  • Nucleosomes / metabolism
  • Rats
  • Taq Polymerase

Substances

  • Nucleosomes
  • DNA
  • Taq Polymerase
  • DNA-Directed DNA Polymerase