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Labeling DNA Probes

Labeling 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...

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Split Hybridization Probe Utilizing a DNA Fluorescent Light-up Aptamer as a Signal Reporter for Sequence-Specific Nucleic Acid Analysis

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

Label-free fluorescent assays based on aptamer-target recognition.

Ying Tan¹, Xin Zhang, Yonghua Xie

¹Guangdong Provincial Key Laboratory of Chemical Biology, Graduate School at Shenzhen, Tsinghua University, Shenzhen, 518055, P. R. China.

|March 28, 2012

Summary

New label-free fluorescent assays utilize DNA hybridization competition for sensitive and selective detection of targets like adenosine triphosphate (ATP) and thrombin. This general method offers a robust approach for molecular sensing applications.

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Area of Science:

Biochemistry
Molecular Biology
Analytical Chemistry

Background:

Developing sensitive and selective assays is crucial for molecular target detection.
Label-free detection methods reduce assay complexity and cost.
Competition between intramolecular DNA hybridization and aptamer-target binding offers a novel sensing mechanism.

Purpose of the Study:

To develop a general, label-free fluorescent assay platform.
To demonstrate the assay's sensitivity and selectivity using model targets.
To validate the competition-based mechanism for aptamer-target binding detection.

Main Methods:

Designed DNA probes capable of intramolecular hybridization.
Exploited the displacement of hybridization upon aptamer-target interaction.
Utilized fluorescence signal changes to quantify target binding.
Tested the assay with adenosine triphosphate (ATP) and thrombin.

Main Results:

Achieved label-free detection of ATP and thrombin.
Demonstrated high sensitivity with low detection limits.
Confirmed high selectivity against non-target molecules.
Validated the assay's general applicability across different targets.

Conclusions:

The developed fluorescent assay is a versatile and effective platform for label-free molecular detection.
The competition between DNA hybridization and aptamer-target binding provides a robust sensing strategy.
This method holds promise for various applications in diagnostics and biochemical research.