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Related Concept Videos

In-situ Hybridization02:31

In-situ Hybridization

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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...
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FISH - Fluorescent In-situ Hybridization02:07

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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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Related Experiment Video

Updated: Oct 5, 2025

High Resolution Whole Mount In Situ Hybridization within Zebrafish Embryos to Study Gene Expression and Function
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In Situ HiC.

Timothy M Johanson1,2, Rhys S Allan3,4

  • 1The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia. johanson@wehi.edu.au.

Methods in Molecular Biology (Clifton, N.J.)
|February 1, 2022
PubMed
Summary
This summary is machine-generated.

In situ HiC is a method to map genome 3D architecture using DNA ligation events. This study details the steps for capturing, sequencing, and analyzing these proximity ligation events.

Keywords:
Chromatin architectureChromosome conformation captureGenome organizationIn situ HiC

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Area of Science:

  • Genomics
  • Molecular Biology
  • Biotechnology

Background:

  • Understanding genome 3D architecture is crucial for gene regulation.
  • Current methods require detailed protocols for accurate reconstruction.

Purpose of the Study:

  • To provide a detailed description of the in situ HiC method.
  • To enable researchers to accurately reconstruct genome 3D architecture.

Main Methods:

  • Utilizing proximity ligation to capture DNA-DNA interactions within fixed nuclei.
  • Employing biotinylated nucleotides for tagging restriction enzyme-digested DNA ends.
  • Next-generation sequencing to determine linear and 3D genomic positions.

Main Results:

  • Successfully captured and sequenced DNA-DNA ligation events.
  • Demonstrated the ability to determine both linear and 3D genomic relationships.

Conclusions:

  • The described in situ HiC protocol facilitates detailed reconstruction of genome 3D architecture.
  • This method is essential for studying genome organization and function.