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Related Concept Videos

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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.
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Label-free protein recognition using aptamer-based fluorescence assay.

Yan Jin1, Jinyan Bai, Hongyan Li

  • 1Key Laboratory of Applied Surface and Colloid Chemistry, Ministry of Education, Key Laboratory of Analytical Chemistry for Life Science of Shaanxi Province, School of Chemistry and Materials Science, Shaanxi Normal University, Xi'an, 710062, China. jinyan@snnu.edu.cn

The Analyst
|May 15, 2010
PubMed
Summary
This summary is machine-generated.

This study introduces a label-free method for detecting thrombin using aptamers and crystal violet. The assay offers ultrasensitive and selective monitoring of thrombin in homogeneous solutions.

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Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Molecular Diagnostics

Background:

  • Real-time protein monitoring in homogeneous solutions without external labels is challenging.
  • Existing methods may require labeling, potentially affecting protein binding properties.

Purpose of the Study:

  • To develop a label-free, ultrasensitive method for detecting thrombin in homogeneous solutions.
  • To utilize aptamers and a fluorescent probe for selective thrombin detection.

Main Methods:

  • Employed a high-affinity thrombin-binding aptamer (TBA) as the molecular recognition probe.
  • Used crystal violet (CV) as the fluorescence signal probe.
  • Monitored fluorescence changes in CV upon TBA-thrombin interaction; circular dichroism (CD) spectra were used to investigate conformational changes.

Main Results:

  • Significant fluorescence enhancement of CV was observed with single-stranded TBA.
  • Thrombin presence caused a decrease in CV fluorescence due to specific TBA-thrombin interaction.
  • Achieved a dynamic response range of 2 x 10(-11) to 2 x 10(-9) M with a detection limit of 8 x 10(-12) M.

Conclusions:

  • The developed method is simple, effective, and label-free for both probe and target.
  • The assay demonstrates high selectivity and ultrasensitivity for thrombin detection.
  • The procedure minimally affects protein binding properties, enabling reliable analysis.