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Updated: Jun 4, 2026
11:36
Simple Method for Fluorescence DNA In Situ Hybridization to Squashed Chromosomes
Published on: January 6, 2015
A simple DNA Characterization method using fiber-fluorescence in situ hybridization performed without DNA
Tamaki Hirose1, Shigeru Sugiyama
1Nanobiotechnology Laboratory, National Food Research Institute, National Agriculture and Food Research Organization, Tsukuba, Ibaraki, Japan.
Photochemistry and Photobiology
|March 4, 2011
Summary
This study introduces a new fluorescence imaging method for visualizing DNA. The technique allows for rapid, direct visualization of specific binding sites on intact DNA molecules without causing damage.
Area of Science:
- Molecular Biology
- Genetics
- Biophysics
Background:
- Traditional fiber-fluorescence in situ hybridization (fiber-FISH) can cause DNA fragmentation.
- Accurate visualization of DNA probe binding sites is crucial for genetic studies.
Purpose of the Study:
- To develop a novel high-resolution fluorescence imaging technique for DNA.
- To enable rapid and direct visualization of specific binding sites on intact DNA molecules.
Main Methods:
- High-resolution fluorescence imaging of lambda phage DNA.
- Hybridization with fluorescent-labeled DNA and peptide nucleic acid probes.
- Liquid hybridization, rapid denaturation, molecular combing, and glass substrate spotting.
Main Results:
- Continuous fluorescence signal observed for intact lambda DNA.
- Probes bound to predicted sites, but with a distinct pattern compared to fiber-FISH.
- Method avoids DNA damage and fluorescence signal fragmentation.
Conclusions:
- The new method allows for rapid and direct visualization of specific binding sites on intact DNA.
- This technique offers an improvement over conventional fiber-FISH by preserving DNA integrity.
- The method is a valuable tool for genetics research and large-scale DNA sequencing projects.
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02:07FISH - 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,...
02:31In-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...
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...