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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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In Situ Hybridization for the Precise Localization of Transcripts in Plants
12:15

In Situ Hybridization for the Precise Localization of Transcripts in Plants

Published on: November 23, 2011

In situ hybridization.

Chung-Jui Tsai1, Scott A Harding

  • 1Warnell School of Forestry and Natural Resources, and Department of Genetics, University of Georgia, GA, USA.

Methods in Cell Biology
|January 16, 2013
PubMed
Summary
This summary is machine-generated.

This study presents a non-radioactive in situ hybridization method for precisely locating messenger RNA (mRNA) transcripts within tissue sections. This technique offers high cellular-level spatial resolution for gene expression analysis.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • Northern hybridization and PCR methods lack cellular-level spatial resolution for RNA analysis.
  • Accurate localization of mRNA transcripts is crucial for understanding gene function and regulation.
  • Existing methods struggle to differentiate between closely related gene family members.

Purpose of the Study:

  • To present a non-radioactive in situ hybridization protocol for mRNA localization in thin tissue sections.
  • To enable high-resolution spatial analysis of transcript distribution at the cellular level.
  • To facilitate the discrimination of closely related gene family members.

Main Methods:

  • Development of a non-radioactive in situ hybridization protocol.
  • Utilization of gene-specific probes for targeted mRNA detection.
  • Application to thin tissue sections for cellular-level analysis.

Main Results:

  • The protocol successfully localizes mRNA transcripts within thin tissue sections.
  • Achieved spatial resolution at the cellular level surpasses Northern hybridization and PCR-based methods.
  • Demonstrated ability to discriminate between closely related gene family members using specific probes.

Conclusions:

  • The non-radioactive in situ hybridization protocol provides precise spatial resolution of mRNA transcripts.
  • This method enhances the study of gene expression patterns at the cellular level.
  • The protocol is scalable for comparative analysis of multiple genes, despite lengthy tissue preparation.