Related Experiment Video
Updated: Jun 6, 2026

In vitro Assembly of Semi-artificial Molecular Machine and its Use for Detection of DNA Damage
Published on: January 11, 2012
In situ ligation simplified: using PCR fragments for detection of double-strand DNA breaks in tissue sections
1Departments of Neurosurgery and Molecular & Cellular Biology, Baylor College of Medicine, and Michael E DeBakey VA Medical Center, Houston, TX, USA. vdidenko@bcm.edu
Abstract:
The simplified in situ ligation procedure is described. All reagents for the assay can be easily obtained in any molecular or cell biology laboratory. The technique uses ligation of double-stranded, PCR-derived DNA fragments labeled with digoxigenin or fluorophores for highly selective detection of apoptotic cells in paraffin-embedded tissue sections. Two types of DNA fragments prepared by PCR are employed. The fragment synthesized by Taq polymerase contains single-base 3' overhangs, whereas the Pfu polymerase-made fragment is blunt ended. Both fragments can be used as specific, sensitive and cost-effective DNA damage probes. After ligation to apoptotic nuclei in tissue sections, they indicate the presence of double-strand DNA breaks with single-base 3' overhangs as well as blunt ends.
More Related Videos
Related Concept Videos
In-situ Hybridization
Types of probes and labels
A probe is a complementary strand of DNA or RNA that binds to corresponding nucleotide sequences in a cell. Many...
DNA Isolation
Southern Blot
Denatured DNA fragments must be transferred onto a carrier membrane from the gel to make it accessible to a probe - a small ssDNA fragment complementary to the target DNA...

