Related Experiment Video
Updated: Jun 23, 2026
07:10
Split Hybridization Probe Utilizing a DNA Fluorescent Light-up Aptamer as a Signal Reporter for Sequence-Specific Nucleic Acid Analysis
Published on: July 8, 2025
Fluorescence probe sensitive to detect G-rich target strands through quenching
Arvind Misra1, Pratibha Dwivedi, Mohammad Shahid
1Nucleic Acids Research Laboratory, Department of Chemistry, Faculty of Science, Banaras Hindu University, 221005 Varanasi, India. arvindmisra2003@yahoo.com
Bioorganicheskaia Khimiia
|April 21, 2009
Summary
A novel modified fluorescent probe, U(F)AA, was developed for DNA diagnostics. Its ability to detect specific DNA sequences through fluorescence changes shows promise for homogeneous hybridization assays.
Area of Science:
- Biochemistry
- Molecular Biology
- Nucleic Acid Chemistry
Background:
- Development of sensitive and specific probes is crucial for DNA diagnostics.
- Hybridization assays require probes that can accurately detect target DNA sequences.
- Fluorescent probes offer advantages in real-time monitoring and high sensitivity.
Purpose of the Study:
- To synthesize and characterize a modified fluorescent probe, U(F)AA.
- To evaluate the probe's performance in hybridization studies with complementary DNA and DNA containing G-rich telomeric units.
- To assess the potential application of this probe in DNA diagnostics.
Main Methods:
- Synthesis of a modified uridine nucleoside analogue incorporating a dansyl group.
- Hybridization studies using the synthesized probe with perfectly matched (U/A) DNA and DNA with G-rich telomeric units.
- Analysis of thermal stability and fluorescence intensity changes upon hybridization.
Main Results:
- Successful synthesis of the modified fluorescent probe U(F)AA.
- Demonstrated hybridization with complementary DNA sequences.
- Observed a decrease in fluorescence intensity in the presence of G-rich telomeric units.
- Thermal stability data indicated probe performance.
Conclusions:
- The modified fluorescent probe U(F)AA shows potential for DNA diagnostics.
- The observed fluorescence quenching by G-rich units highlights its utility in homogeneous hybridization assays.
- This approach offers a sensitive method for detecting specific DNA sequences.
Related Concept Videos
03:31Labeling DNA Probes
DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...
02:11Reporter Genes
Reporter genes are a type of protein-coding gene that are often tagged to a gene of interest. Once inside a target cell, reporter genes usually produce visually identifiable characteristics like fluorescence and luminescence when expressed along with the gene of interest. Thus, reporter genes “report” the presence or absence of genes of interest in an organism, determine the gene expression pattern, or track the physical location of a DNA segment or protein in the cell.
Commonly used reporter...
Commonly used reporter...
02:07FISH - Fluorescent In-situ Hybridization
Fluorescence in situ hybridization, or FISH, was developed in the early 1980s and has quickly become one of the most widely used techniques in cytogenetics. Labeled probes are used to bind complementary DNA or RNA sequences on a chromosome or in a region within a cell. Earlier, the probes could only be obtained by cloning or reverse transcription of a DNA template. Currently, the probe oligonucleotides can be synthesized synthetically. Additionally, with the advancement of optical techniques,...
01:19Protein Dynamics in Living Cells
Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...