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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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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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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...
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In Situ Hybridization for the Precise Localization of Transcripts in Plants
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Technical review: In situ hybridization.

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In situ hybridization detects DNA or RNA sequences in tissues using labeled probes. This versatile technique has broad research and diagnostic applications.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Cell Biology

Background:

  • In situ hybridization (ISH) is a powerful cytogenetic technique.
  • It visualizes the spatial distribution of specific DNA or RNA sequences within cells or tissues.
  • ISH relies on the principle of nucleic acid hybridization.

Purpose of the Study:

  • To review the fundamental principles of in situ hybridization.
  • To discuss various types of ISH, their applications, and limitations.
  • To highlight the utility of ISH in research and diagnostics.

Main Methods:

  • Utilizes labeled nucleotide probes (radio-, fluorescent-, or antigen-labeled) that bind to complementary target DNA or RNA sequences.
  • Detection methods include autoradiography, fluorescence microscopy, and immunohistochemistry, depending on the probe label.
  • The standard ISH process involves probe hybridization and subsequent visualization.

Main Results:

  • ISH enables precise localization of nucleic acid targets within their native cellular or tissue context.
  • Different ISH variations offer distinct advantages for specific applications.
  • The technique is adaptable for various detection systems.

Conclusions:

  • In situ hybridization is an indispensable tool in biological and medical research.
  • Its diagnostic potential, particularly in identifying genetic alterations or pathogen presence, is significant.
  • Understanding ISH variations is crucial for optimizing experimental design and interpretation.