Dna Lesion Quantification
Dna Lesion Quantification - Related Videos
10:59
Atomic Force Microscopy Investigations of DNA Lesion Recognition in Nucleotide Excision Repair
0 Views •
Cited by 2 •
2017
Here, the study of different DNA lesion recognition approaches via single molecule AFM imaging is demonstrated with the nucleotide excision repair system as an example. The procedures of DNA and protein sample preparations and experimental as well as analytical details for the AFM experiments are described.
12:43
Advanced Confocal Microscopy Techniques to Study Protein-protein Interactions and Kinetics at DNA Lesions
0 Views •
Cited by 8 •
2017
Laser microirradiation is a useful tool for studies of DNA repair in living cells. A methodological approach for the use of UVA lasers to induce various DNA lesions is shown. We have optimized a method for local microirradiation that maintains the normal cell cycle; thus, irradiated cells proceed through mitosis.
12:15
Quantification of three DNA Lesions by Mass Spectrometry and Assessment of Their Levels in Tissues of Mice Exposed to Ambient Fine Particulate Matter
0 Views •
Cited by 1 •
2019
We describe here methods for sensitive and accurate quantification of the lesions 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodGuo), 1,N6-etheno-2'-deoxyadenosine (1,N6-dAdo) and 1,N2-etheno-2'-deoxyguanosine (1,N2-dGuo) in DNA. The methods were applied to the assessment of the effects of ambient fine particulate matter (PM2.5) in tissues (lung, liver and kidney) of exposed A/J mice.
08:16
Comparative Lesions Analysis Through a Targeted Sequencing Approach
0 Views •
2019
This article describes a method to identify clonal and subclonal alterations among different specimens from a given patient. Although the experiments described here focus on a specific tumor type, the approach is broadly applicable to other solid tumors.
02:10
Translesion DNA Polymerases
0 Views •
2020
Translesion (TLS) polymerases rescue stalled DNA polymerases at sites of damaged bases by replacing the replicative polymerase and installing a nucleotide across the damaged site. Doing so, TLS allows additional time for the cell to repair the damage before resuming regular DNA replication. TLS polymerases are found in all three domains of life - archaea, bacteria, and eukaryotes. Of the different classes of TLS polymerases, members of the Y family are fitted with specialized structures that...
FAQs
The assay links lesion abundance to a detectable change, such as altered electrophoretic migration, antibody binding, enzymatic conversion, or a mass spectrometric signal. Results are interpreted relative to undamaged DNA or a defined standard, allowing researchers to distinguish greater or lesser genome damage rather than relying only on qualitative observations.
Different lesions present different chemical or structural features, so no single detection strategy represents all forms of damage equally. Strand breaks may be assessed through migration behavior, whereas base modifications or DNA adducts may require lesion-specific antibodies, enzymatic conversion, or mass spectrometry. Method selection therefore determines which component of genome integrity is measured.
The biological source of the analyzed DNA determines whose genome the measurement reflects, while the selected assay determines which lesion class is visible. In infection studies, researchers can therefore examine damage associated with host responses, pathogen-associated processes, or both, provided the experimental design distinguishes the relevant DNA populations and compares their signals appropriately.
A useful workflow begins by identifying the lesion class and deciding whether the experiment requires migration-based, antibody-based, enzymatic, or mass spectrometric detection. DNA is then evaluated against undamaged material or a defined standard, and the resulting signal is interpreted as relative lesion burden. This sequence connects assay choice with a specific genome-integrity question.
It can reveal how pathogens, inflammatory oxidants, antimicrobial responses, or therapeutic compounds affect host and microbial genomes. Comparing lesion measurements across these conditions helps investigators determine whether genome damage accompanies infection or treatment and provides a quantitative basis for examining infection-related genotoxicity rather than inferring it from exposure alone.
Changes in lesion burden can help clarify mechanisms of infection-related genotoxicity and assess how DNA repair relates to experimental conditions. The measurements also support studies of disease progression and treatment effects by providing a genome-integrity outcome that can be compared with pathogen exposure, inflammatory or antimicrobial activity, and therapeutic intervention.