Dna Synthesis Assay
Dna Synthesis Assay - Related Videos
02:54
DNA Synthesis Assay: A Technique to Assess DNA Synthesis in Proliferating Cells Using Radioactive Tritiated Thymidine Incorporation
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2023
In this video, we determine the proliferative capacity of neuronal progenitor cells by measuring the incorporation of radioactive tracer tritiated thymidine into the DNA of cells. This technique can be used to study cell proliferation after treatment with certain drugs and growth factors.
10:00
Assaying DNA Damage in Hippocampal Neurons Using the Comet Assay
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Cited by 21 •
2012
The comet assay is an efficient way of detecting single- and double-strand breaks, including alkali-labile sites and DNA-DNA/DNA-protein cross-links on the DNA in all cells including hippocampal neurons. The method takes advantage of the differential migration of DNA in an electric field due to differences in amount of DNA damage.
01:28
Inhibitors of Bacterial DNA Synthesis
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2026
Bacterial pathogens depend on precise and efficient DNA replication to sustain infection. Two type II topoisomerases—DNA gyrase and topoisomerase IV—are critical to this process, as they resolve DNA supercoiling and unlink chromosomes during replication. Fluoroquinolones, synthetic derivatives of quinolones, exploit this mechanism by stabilizing the transient DNA–enzyme cleavage complex, preventing strand religation, and causing lethal double-strand breaks. These antibiotics are selectively...
07:38
DNA Polymerase Activity Assay Using Near-infrared Fluorescent Labeled DNA Visualized by Acrylamide Gel Electrophoresis
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Cited by 2 •
2017
This protocol describes the characterization of DNA polymerase synthesis of modified DNA through observation of changes to near-infrared fluorescently labeled DNA using gel electrophoresis and gel imaging. Acrylamide gels are used for high resolution imaging of the separation of short nucleic acids, which migrate at different rates depending on size.
08:14
Kinetics of Lagging-strand DNA Synthesis In Vitro by the Bacteriophage T7 Replication Proteins
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Cited by 3 •
2017
We describe sensitive, gel-based discontinuous assays to examine the kinetics of lagging-strand initiation using the replication proteins of bacteriophage T7.
FAQs
The primer supplies the starting point for extension, while the template determines which complementary deoxynucleotides can be incorporated. This pairing connects the measured signal to polymerase activity rather than to DNA quantity alone. Changing the primer-template arrangement can therefore help investigators examine how effectively a polymerase performs a defined synthesis reaction.
A DNA synthesis assay can report activity through labeled nucleotides, fluorescent products, or amplified DNA. These readouts differ in how newly produced DNA becomes detectable, but each provides a quantitative measurement of synthesis. The selected format can be matched to the experimental goal, such as comparing polymerase activity or evaluating an inhibitor.
By measuring how much DNA is produced, investigators can characterize polymerase kinetics, meaning the behavior of synthesis under the assay conditions. Comparing synthesis in the presence and absence of a compound can show whether that compound inhibits or alters activity. This makes the assay useful for connecting biochemical changes with polymerase function.
A basic workflow combines a template strand, a primer, DNA polymerase, and complementary deoxynucleotides, followed by detection of the newly synthesized DNA. The primer is extended on the template, and the resulting product is quantified using a label, fluorescence, or amplification-based signal. This sequence links reaction setup directly to the final measurement.
Researchers use this assay when they need a quantitative view of DNA polymerase activity, genome replication, or DNA repair pathways. It can also support compound testing by revealing changes in synthesis. In biochemistry, the method is especially useful for examining enzyme behavior in a controlled reaction and comparing how different experimental conditions affect DNA production.
Measurements from these assays can help determine whether DNA synthesis is functioning normally or has been altered by repair-related processes or inhibitory compounds. Such results contribute to studies of genome stability, because replication and repair pathways depend on controlled DNA production. The assay therefore connects a biochemical measurement with broader questions about how genomes are maintained.