Related Experiment Videos
A quantitative assay for fragmented DNA in apoptotic cells
1Department of Medical Oncology, University of Texas M. D. Anderson Cancer Center, Houston 77030.
Analytical Biochemistry
|November 15, 1992
Summary
Researchers developed a sensitive assay to quantify DNA fragmentation in apoptotic cells. This method detects picogram levels of DNA damage, aiding in the study of programmed cell death.
Area of Science:
- Molecular Biology
- Cell Biology
- Biochemistry
Background:
- Internucleosomal DNA fragmentation is a hallmark of apoptosis.
- Quantifying DNA fragmentation is crucial for understanding apoptosis.
- Existing methods lack sufficient sensitivity.
Purpose of the Study:
- To develop a highly sensitive assay for quantifying internucleosomal DNA fragmentation in apoptotic cells.
- To enable the detection of DNA fragments at picogram levels.
Main Methods:
- Purification of cellular DNA.
- Dephosphorylation of DNA ends.
- Radiolabeling of DNA with 32P.
- Agarose gel electrophoresis.
- Quantitation of radioactivity in DNA bands.
Main Results:
- The assay is 1000- to 2000-fold more sensitive than ethidium bromide staining.
- Detection of DNA fragments at picogram levels is achievable.
- The assay can quantitatively determine the number of fragmented DNA strands.
Conclusions:
- A novel, highly sensitive assay for DNA fragmentation in apoptosis was developed.
- This assay facilitates detailed studies of the time course of DNA fragmentation.
- The method is applicable to cells undergoing apoptosis induced by various agents.