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Related Concept Videos

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,...
Labeling DNA Probes03:31

Labeling DNA Probes

DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...
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...
Southern Blot02:57

Southern Blot

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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Related Experiment Video

Updated: May 27, 2026

Robust 3D DNA FISH Using Directly Labeled Probes
12:16

Robust 3D DNA FISH Using Directly Labeled Probes

Published on: August 15, 2013

A quick and simple FISH protocol with hybridization-sensitive fluorescent linear oligodeoxynucleotide probes.

Dan Ohtan Wang1, Hitomi Matsuno, Shuji Ikeda

  • 1Institute for Integrated Cell-Material Sciences, Kyoto University, Kyoto 606-8501, Japan. dwang@icems.kyoto-u.ac.jp

RNA (New York, N.Y.)
|November 22, 2011
PubMed
Summary

A new fluorescence in situ hybridization (FISH) method, ECHO-FISH, offers rapid, high-resolution detection of DNA and RNA. This simplified protocol accelerates cytogenetic and gene-expression analysis without compromising accuracy.

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Split Hybridization Probe Utilizing a DNA Fluorescent Light-up Aptamer as a Signal Reporter for Sequence-Specific Nucleic Acid Analysis
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Split Hybridization Probe Utilizing a DNA Fluorescent Light-up Aptamer as a Signal Reporter for Sequence-Specific Nucleic Acid Analysis

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Area of Science:

  • Molecular Biology
  • Genetics
  • Cell Biology

Background:

  • Fluorescence in situ hybridization (FISH) is a vital technique for genetic analysis, including karyotyping, cancer diagnosis, and gene expression studies.
  • Conventional FISH protocols are often lengthy and complex, posing limitations for rapid analysis and high-throughput applications.

Purpose of the Study:

  • To develop a streamlined and efficient FISH method.
  • To enable rapid, high-resolution detection of both DNA and RNA sequences.
  • To overcome the limitations of conventional FISH protocols.

Main Methods:

  • Development of a novel FISH method utilizing exciton-controlled hybridization-sensitive fluorescent oligodeoxynucleotide (ECHO) probes.
  • Implementation of a rapid 25-minute protocol from sample fixation to mounting, eliminating stringency washing steps.
  • Application of ECHO-FISH for multicolor detection of specific DNA and RNA sequences.

Main Results:

  • ECHO-FISH successfully detected specific DNA and RNA sequences with high resolution and reproducibility.
  • The method enabled visualization of intranuclear poly(A) RNA speckles and dendritic mRNAs in neuronal cultures.
  • Detection of telomeric and centromeric DNA on metaphase chromosomes was demonstrated.

Conclusions:

  • ECHO-FISH provides a significantly simplified and accelerated alternative to conventional FISH.
  • The method is highly stringent, reproducible, and compatible with other cellular labeling techniques.
  • ECHO-FISH is poised to advance cytogenetic and gene-expression analysis due to its speed and high resolution.