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,metabolism
DNA Fragmentation
DNA-Directed DNA Polymerase/metabolism
Electrophoresis, Agar Gel/methods
Nucleosomes/metabolism
Rats
Taq Polymerase
Chemicals
Nucleosomes
DNA
Taq Polymerase
DNA-Directed DNA Polymerase
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Eldadah B A
Department of Neurology and the Georgetown Institute for Cognitive and Computational Sciences, Georgetown University Medical Center, Washington, DC 20007, USA.
Yakovlev A G
Faden A I
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