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Related Concept Videos

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 Polymerase Activity Assay Using Near-infrared Fluorescent Labeled DNA Visualized by Acrylamide Gel Electrophoresis
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Visual detection of labeled oligonucleotides using visible-light-polymerization-based amplification.

Ryan R Hansen1, Hadley D Sikes, Christopher N Bowman

  • 1Department of Chemical and Biological Engineering, ECCH 111 CB 424, University of Colorado, Boulder, Colorado 80309, USA.

Biomacromolecules
|December 7, 2007
PubMed
Summary

This study presents a rapid, nonenzymatic DNA biochip method for sensitive genetic diagnostics. It achieves significant signal amplification using photopolymerization, enabling detection with inexpensive equipment.

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Area of Science:

  • Biotechnology
  • Molecular Diagnostics
  • Bioengineering

Background:

  • DNA biochips offer potential for rapid genetic screening.
  • Current limitations include simultaneous identification of low copy number targets with inexpensive detection.

Purpose of the Study:

  • To develop a rapid, nonenzymatic signal amplification method for DNA biochips.
  • To enable sensitive detection of low copy number polynucleotides using affordable instrumentation.

Main Methods:

  • A surface-initiated photopolymerization technique was employed on a glass microarray.
  • Visible light photoinitiators coupled to streptavidin bound biotin-labeled DNA capture sequences.
  • Hydrogel layers were formed via photopolymerization upon exposure to monomers and light.

Main Results:

  • Achieved a 10(6) to 10(7) amplification factor.
  • Enabled detection of approximately 10(4) labeled oligonucleotides.
  • Demonstrated a rapid (20 min) assay with minimal instrumentation (optical microscope/CCD camera).

Conclusions:

  • The presented method offers a rapid and sensitive approach for genetic diagnostics on biochips.
  • Nonenzymatic photopolymerization provides a viable signal amplification strategy for low-cost detection.
  • This technology addresses limitations in current DNA biochip diagnostic methodologies.