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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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Following Cell-fate in E. coli After Infection by Phage Lambda
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Published on: October 14, 2011

Quantitative characterization of quantum dot-labeled lambda phage for Escherichia coli detection.

Peter B Yim1, Matthew L Clarke, Michael McKinstry

  • 1National Institute of Standards and Technology, Gaithersburg, Maryland 20899, USA.

Biotechnology and Bioengineering
|July 28, 2009
PubMed
Summary

Genetically modified bacteriophage were decorated with quantum dots (QDs) for bacterial detection. This method offers a sensitive approach for identifying bacterial cells using fluorescent nanocrystal-biological conjugates.

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Area of Science:

  • Biotechnology
  • Nanotechnology
  • Microbiology

Background:

  • Quantum dots (QDs) offer unique optical properties for bio-imaging.
  • Bacteriophage can be engineered for specific biological detection applications.
  • Developing robust methods for conjugating nanoparticles to biological entities is crucial.

Purpose of the Study:

  • To characterize the binding of Cadmium Selenide/Zinc Sulfide (CdSe/ZnS) quantum dots to genetically modified bacteriophage.
  • To establish a model system for bacterial detection using QD-decorated bacteriophage.
  • To quantify QD attachment to bacteriophage capsids and compare binding efficiency between different phage types.

Main Methods:

  • Flow cytometry and image-based cytometry for analyzing fluorescently labeled bacteria and phage binding.
  • Transmission electron microscopy (TEM) for visualizing QD-phage conjugates.
  • Electrospray differential mobility analysis (EDMA) for quantifying QD-phage binding.

Main Results:

  • Quantified 4-17 QDs attached per lambda phage capsid.
  • Demonstrated significantly higher QD binding to lambda phage compared to bacteriophage T7 (nearly four-fold increase).
  • Image-based cytometry confirmed fluorescent labeling of bacteria and quantified decorated phage bound to cells.

Conclusions:

  • Genetically modified bacteriophage serve as an effective platform for QD conjugation for bacterial detection.
  • The developed characterization methodology is applicable to various fluorescent nanocrystal-biological conjugates.
  • This approach enhances sensitivity and specificity in bacterial detection systems.