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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...
Southern Blot02:57

Southern Blot

Agarose gel electrophoresis is very useful in separating DNA fragments by size. Running a DNA ladder containing fragments of the known length alongside the sample helps determine the approximate length of the sample DNA fragments. However, additional steps are needed to verify the sequence identity of the sample DNA fragments.
Denatured DNA fragments must be transferred onto a carrier membrane from the gel to make it accessible to a probe - a small ssDNA fragment complementary to the target DNA...
In-situ Hybridization02:31

In-situ Hybridization

In situ hybridization (ISH) is a technique used to detect and localize specific DNA or RNA molecules in cells, tissue, or tissue sections using a labeled probe. The technique was first used in 1969 for the investigation of nucleic acids. It is currently an essential tool in scientific research and clinical settings, especially for diagnostic purposes.
Types of probes and labels
A probe is a complementary strand of DNA or RNA that binds to corresponding nucleotide sequences in a cell. Many...
Sanger Sequencing01:57

Sanger Sequencing

DNA sequencing is a fundamental technique that is routinely used in the biological sciences. This method can be applied to a range of questions at different scales - from the sequencing of a cloned DNA fragment or the study of a mutation in a gene up to whole-genome sequencing. However, despite the widespread use of sequencing today, it was not until 1977 that Fredrick Sanger and his collaborators developed the chain-termination method to decode DNA sequences. It relies on the separation of a...
DNA Microarrays02:34

DNA Microarrays

Microarrays are high-throughput and relatively inexpensive assays that can be automated to analyze large quantities of data at a time. They are used in genome-wide studies to compare gene or protein expression under two varied conditions, such as healthy and diseased states. Microarrays consist of glass or silica slides on which probe molecules are covalently attached through surface functionalization. Most commonly, the slides are prepared through the chemisorption of silanes to silica...
Western Blotting01:15

Western Blotting

Western blotting is an analytical technique for protein identification. It has various applications in immunology and medicine, including detecting diseases like bovine spongiform encephalopathy, mad cow disease, and human and feline immunodeficiency virus from biological samples.
The technique begins with separating proteins from the sample using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), followed by protein transfer, immunoblotting, and finally, protein detection.

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Related Experiment Video

Updated: Jun 3, 2026

In Vitro Biochemical Assays using Biotin Labels to Study Protein-Nucleic Acid Interactions
08:14

In Vitro Biochemical Assays using Biotin Labels to Study Protein-Nucleic Acid Interactions

Published on: July 17, 2019

Detection of DNA sequences using biotinylated probes.

J L Woodhead1, H Figueiredo, A D Malcolm

  • 1Department of Biochemistry, Charing Cross and Westminster Medical School, London, UK.

Methods in Molecular Biology (Clifton, N.J.)
|March 23, 2011
PubMed
Summary

Biotinylated deoxyuridine triphosphate (dUTP) analogs serve as effective substrates for DNA polymerase. This enables the creation of biotin-labeled DNA for detection using methods like streptavidin-peroxidase.

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Area of Science:

  • Molecular Biology
  • Biochemistry
  • Nucleic Acid Chemistry

Background:

  • Biotinylation is a crucial technique for labeling and detecting biomolecules.
  • Deoxyuridine triphosphate (dUTP) analogs are essential building blocks in molecular biology.
  • Developing efficient methods for DNA labeling is vital for various biological assays.

Purpose of the Study:

  • To investigate the utility of biotinylated dUTP analogs as substrates for DNA polymerase.
  • To demonstrate the incorporation of biotin into DNA using these analogs.
  • To establish a method for detecting biotin-labeled DNA.

Main Methods:

  • Synthesis of biotinylated dUTP analogs with an allylamine linker at the C-5 position.
  • Enzymatic incorporation of biotinylated dUTP into DNA using DNA polymerase.
  • Hybridization of biotin-labeled DNA to target sequences.
  • Detection of biotin probes using streptavidin-peroxidase conjugates.

Main Results:

  • Biotinylated dUTP analogs were successfully utilized as substrates for DNA polymerase.
  • DNA molecules were efficiently biotinylated at the C-5 position of the pyrimidine ring.
  • The resulting biotin-substituted DNA retained hybridization properties similar to unsubstituted DNA.
  • Detection of the incorporated biotin probe was achieved using streptavidin-peroxidase.

Conclusions:

  • Biotinylated dUTP analogs are valuable tools for DNA labeling.
  • This method provides a reliable way to generate detectable DNA probes.
  • The technique facilitates the localization of specific DNA sequences through biotin-streptavidin interactions.