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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.
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Fluorescence in situ hybridization, or FISH, was developed in the early 1980s and has quickly become one of the most widely used techniques in cytogenetics. Labeled probes are used to bind complementary DNA or RNA sequences on a chromosome or in a region within a cell. Earlier, the probes could only be obtained by cloning or reverse transcription of a DNA template. Currently, the probe oligonucleotides can be synthesized synthetically. Additionally, with the advancement of optical techniques,...
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Real Time RT-PCR

Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
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Detection of Bacteria Using Fluorogenic DNAzymes
13:20

Detection of Bacteria Using Fluorogenic DNAzymes

Published on: May 28, 2012

A fluorogenic, nucleic acid directed "click" reaction.

Elmar Jentzsch1, Andriy Mokhir

  • 1Anorganisch-Chemisches Institut, Ruprecht-Karls-Universität Heidelberg, Im Neuenheimer Feld 270, 69120 Heidelberg, Germany.

Inorganic Chemistry
|September 23, 2009
PubMed
Summary
This summary is machine-generated.

A new copper-catalyzed reaction enables sensitive, sequence-specific detection of nucleic acids by monitoring fluorescence. Even a single mismatch prevents the reaction, highlighting its high specificity for DNA and RNA analysis.

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Chemical Biology

Background:

  • Nucleic acid detection is crucial for diagnostics and research.
  • Existing methods may lack sensitivity or specificity.
  • Developing novel, efficient detection strategies is important.

Purpose of the Study:

  • To develop a novel fluorogenic assay for sequence-specific nucleic acid detection.
  • To investigate the efficiency of copper(I)-catalyzed reactions for this purpose.
  • To assess the impact of single nucleotide mismatches on reaction efficiency.

Main Methods:

  • A copper(I)-catalyzed chemical reaction was designed to be directed by nucleic acid sequences.
  • The reaction's progress was monitored by an increase in fluorescence intensity.
  • The catalytic activity of copper(I) ions stabilized by water-soluble ligands was compared to free copper(I).

Main Results:

  • The assay achieved sequence-specific detection of nucleic acids down to 20 nM concentrations.
  • A single nucleotide mismatch in the template strand completely inhibited the reaction.
  • Copper(I) ions stabilized with water-soluble ligands demonstrated significantly higher catalytic efficiency than free copper(I).

Conclusions:

  • A highly sensitive and specific fluorogenic assay for nucleic acid detection was successfully developed.
  • The assay leverages a copper(I)-catalyzed reaction with high fidelity for sequence recognition.
  • Stabilized copper(I) catalysts offer enhanced performance for nucleic acid detection applications.