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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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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.
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Related Experiment Video

Updated: Nov 10, 2025

In Situ Hybridization for the Precise Localization of Transcripts in Plants
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An RNA in situ hybridization protocol optimized for monocot tissue.

Nora R Zöllner1, Margaret Bezrutczyk1, Reinout Laureyns2,3

  • 1Institute for Molecular Physiology, Heinrich-Heine-University Düsseldorf, Düsseldorf 40225, Germany.

STAR Protocols
|April 2, 2021
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Summary

This study presents an optimized RNA in situ hybridization protocol for maize leaf tissue. The method uses unique antisense probes for accurate gene expression analysis in various plant tissues.

Keywords:
Cell BiologyIn Situ HybridizationMolecular Biology

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Area of Science:

  • Plant biology
  • Molecular biology
  • Genetics

Background:

  • RNA in situ hybridization is a powerful technique for visualizing gene expression within tissues.
  • Traditional RNA in situ hybridization protocols can be lengthy and challenging to optimize.

Purpose of the Study:

  • To provide an optimized and streamlined protocol for RNA in situ hybridization.
  • To enhance the accessibility and troubleshootability of RNA in situ hybridization for plant tissues.

Main Methods:

  • Development of an optimized protocol for RNA in situ hybridization.
  • Generation of three unique antisense probes (>100 bp) per gene of interest.
  • Hybridization of probes to tissue sections of maize leaf, shoot apical meristems, embryos, and floral organs.

Main Results:

  • A refined protocol for RNA in situ hybridization applicable to diverse plant tissues.
  • Demonstrated utility in maize leaf tissue, with potential for broader plant science applications.
  • Successful generation and application of unique antisense probes for gene expression analysis.

Conclusions:

  • The optimized protocol simplifies RNA in situ hybridization, making it more reliable and efficient.
  • This method facilitates detailed spatial analysis of gene expression in plants.
  • The protocol is adaptable for various plant tissues, supporting diverse research needs.