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Labeling DNA Probes03:31

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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[DNA duplex-based fluorescence probes/sensors using monomer-excimer switching].

Kazuhisa Fujimoto1, Masahiko Inouye

  • 1Graduate School of Pharmaceutical Sciences, University of Toyama, Toyama City, Japan. fujimoto@pha.u-toyama.ac.jp

Yakugaku Zasshi : Journal of the Pharmaceutical Society of Japan
|November 5, 2008
PubMed
Summary

This review details DNA-based fluorescence sensors using pyrene excimer-monomer switching. These sensors, including molecular beacons and antibody-based designs, offer sensitive and specific detection of DNA targets and other analytes.

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Area of Science:

  • Molecular biology
  • Biochemistry
  • Analytical chemistry

Context:

  • Development of sensitive and specific fluorescent probes for molecular detection.
  • Exploration of pyrene monomer-excimer switching as a signaling mechanism.
  • Application of DNA structures and antibody motifs in sensor design.

Purpose:

  • To review DNA duplex-based fluorescence probes and sensors utilizing pyrene monomer-excimer switching.
  • To present two main classes: excimer-monomer switching molecular beacons (EMS-MBs) and antibody-based sensors.
  • To highlight the principles and applications of these pyrene-based sensing systems.

Summary:

  • Discusses EMS-MBs with dual pyrene fluorophores on oligonucleotides, enabling unambiguous DNA detection via excimer-to-monomer emission switching upon hybridization.
  • Introduces antibody-motif sensors incorporating guest-binding, DNA, and pyrene sites for detecting specific analytes like potassium ions or porphyrins.
  • Highlights the ability of EMS-MBs to detect 19-mer DNA targets and discriminate single-nucleotide mismatches at 1 nM, and their use in kinetic studies.

Impact:

  • Enables precise detection and discrimination of DNA sequences at low concentrations.
  • Paves the way for novel fluorescent sensors with tailored specificities using antibody-like structures.
  • Expands the toolkit for molecular sensing, with potential applications in diagnostics and biochemical analysis.