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Related Experiment Video

Updated: Apr 10, 2026

Aptamer-Based Target Detection Facilitated by a 3-Stage G-Quadruplex Isothermal Exponential Amplification Reaction
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A G-quadruplex-based Label-free Fluorometric Aptasensor for Adenosine Triphosphate Detection.

Li Juan Li1, Xue Tian, Xiang Juan Kong

  • 1State Key Laboratory of Chemo/Bio-Sensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University.

Analytical Sciences : the International Journal of the Japan Society for Analytical Chemistry
|June 12, 2015
PubMed
Summary

This study presents a label-free fluorescence assay for adenosine triphosphate (ATP) detection using G-quadruplex DNA structures. The assay offers a sensitive method for detecting ATP and potentially other targets with a high signal-to-noise ratio.

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Analytical Chemistry

Background:

  • Adenosine triphosphate (ATP) is a crucial molecule in cellular processes.
  • Developing sensitive and label-free detection methods for ATP is essential for biological research.
  • Existing assays may suffer from high background noise or limited sensitivity.

Purpose of the Study:

  • To develop a novel, label-free fluorescence assay for sensitive adenosine triphosphate (ATP) detection.
  • To utilize G-quadruplex formation for enhanced signal-to-noise ratio in ATP sensing.
  • To demonstrate the versatility of the assay for detecting other biological targets.

Main Methods:

  • A G-quadruplex-based assay utilizing ATP-aptamer hybridized with its complementary strand.
  • Employing SYBR Green I (SG I) as a fluorescent probe and Exonuclease III (Exo III) for dsDNA digestion.
  • Monitoring fluorescence changes upon ATP binding and subsequent G-quadruplex formation.

Main Results:

  • In the absence of ATP, dsDNA is digested by Exo III, yielding low background fluorescence.
  • ATP binding induces aptamer folding into a G-quadruplex, resisting Exo III digestion and leading to high fluorescence.
  • The assay detected ATP in a concentration range of 50 μM to 5 mM with a high signal-to-noise ratio.

Conclusions:

  • A robust and sensitive label-free fluorescence assay for ATP detection was successfully developed.
  • The G-quadruplex-based approach significantly reduces background noise, improving detection sensitivity.
  • The assay demonstrates potential for sensitive determination of various other targets beyond ATP.