Related Experiment Video
Updated: Jun 3, 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
Luminescent detection of specific DNA sequences
J L Woodhead1, H Figueiredo, A D Malcolm
1Department of Biochemistry, Charing Cross and Westminster Medical School, London, UK.
Methods in Molecular Biology (Clifton, N.J.)
|March 23, 2011
Summary
Chemiluminescence occurs when luminol is oxidized, producing light. Adding luciferin enhances this reaction, prolonging light emission for several minutes, measurable with a luminometer or photographic film.
Area of Science:
- Biochemistry
- Chemiluminescence
Background:
- Luminescence is light emission from chemical reactions.
- Oxidation of luminol with hydrogen peroxide and horseradish peroxidase (HRP) produces light.
Purpose of the Study:
- To describe the chemiluminescent reaction of luminol.
- To explain the role of luciferin in enhancing and prolonging light emission.
Main Methods:
- Oxidation of luminol using hydrogen peroxide and HRP as a catalyst.
- Measurement of light emission using a luminometer or photographic film.
Main Results:
- The oxidation reaction produces light.
- Luciferin significantly enhances the reaction intensity.
- Luciferin prolongs light emission for several minutes.
Conclusions:
- The luminol-HRP-hydrogen peroxide system is a source of chemiluminescence.
- Luciferin is an effective enhancer and prolonger of luminol-based light emission.
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...
01:14Photoluminescence: Applications
Photoluminescence offers a wide range of applications due to its inherent sensitivity and selectivity. This technique allows for both direct and indirect analyses of the analyte. Direct quantitative analysis is possible when the analyte exhibits a favorable quantum yield for fluorescence or phosphorescence. However, an indirect analysis may be feasible if the analyte is not fluorescent or phosphorescent, or if the quantum yield is unfavorable. Indirect methods include reacting the analyte with...
02:57Southern Blot
Agarose gel electrophoresis is very useful in separating DNA fragments by size. Running a DNA ladder containing fragments of the known length alongside the sample helps determine the approximate length of the sample DNA fragments. However, additional steps are needed to verify the sequence identity of the sample DNA fragments.
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...
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...
01:57Sanger Sequencing
DNA sequencing is a fundamental technique that is routinely used in the biological sciences. This method can be applied to a range of questions at different scales - from the sequencing of a cloned DNA fragment or the study of a mutation in a gene up to whole-genome sequencing. However, despite the widespread use of sequencing today, it was not until 1977 that Fredrick Sanger and his collaborators developed the chain-termination method to decode DNA sequences. It relies on the separation of a...