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Related Concept Videos

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

FISH - Fluorescent In-situ Hybridization

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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Understanding Early Organogenesis Using a Simplified In Situ Hybridization Protocol in Xenopus
14:50

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Published on: January 12, 2015

Gene expression analysis in developing embryos: in situ hybridization.

Siew-Ging Gong1

  • 1Department of Orthodontics, University of Toronto, Toronto, ON, Canada. sg.gong@dentistry.utoronto.ca

Methods in Molecular Biology (Clifton, N.J.)
|June 7, 2012
PubMed
Summary

In situ hybridization (ISH) allows researchers to visualize gene expression patterns in developing embryos. This method uses nonradioactive riboprobes for precise spatial and temporal analysis of molecular changes.

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Area of Science:

  • Molecular Biology
  • Developmental Biology
  • Toxicology

Background:

  • In situ hybridization (ISH) is crucial for understanding gene expression during development.
  • ISH enables the detection of gene transcript distribution in organisms.
  • It aids in studying molecular changes due to toxicological effects and characterizing novel genes.

Purpose of the Study:

  • To describe a procedure for in situ hybridization (ISH) analysis.
  • To analyze gene transcript spatial distribution in developing whole mouse embryos.
  • To utilize nonradioactive riboprobes for enhanced detection.

Main Methods:

  • The study employed in situ hybridization (ISH) on developing whole mouse embryos.
  • Nonradioactive, in vitro transcribed antisense RNA (riboprobes) were synthesized.
  • Detection of riboprobes was achieved indirectly via a colorimetric reaction.

Main Results:

  • The described procedure allows for the spatial and temporal analysis of gene expression.
  • ISH provides a method to assay for alterations in gene transcript distribution.
  • The technique offers relative ease, speed, and specificity in molecular analysis.

Conclusions:

  • This ISH procedure is an invaluable tool for understanding molecular mechanisms in development.
  • It facilitates hypothesis generation regarding gene function, molecular interactions, and signaling pathways.
  • The nonradioactive ISH method is effective for studying gene expression patterns in embryos.