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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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FISH - Fluorescent In-situ Hybridization02:07

FISH - Fluorescent In-situ Hybridization

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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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Related Experiment Video

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Whole Mount RNA Fluorescent in situ Hybridization of Drosophila Embryos
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Whole Mount RNA Fluorescent in situ Hybridization of Drosophila Embryos

Published on: January 30, 2013

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Simultaneous RNA-DNA FISH.

Lan-Tian Lai1, Zhenyu Meng2, Fangwei Shao3

  • 1School of Biological Sciences, Nanyang Technological University, Singapore, Singapore.

Methods in Molecular Biology (Clifton, N.J.)
|August 21, 2021
PubMed
Summary
This summary is machine-generated.

This study introduces a new RNA-DNA FISH method to simultaneously detect RNA and DNA. The improved protocol protects fragile RNA signals during DNA denaturation, enabling accurate molecular analysis.

Keywords:
Amino-labeled ProbesFluorescence in situ hybridization (FISH)Simultaneous RNA-DNA FISHSingle Molecule RNA FISHlncRNA

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Area of Science:

  • Molecular Biology
  • Cell Biology
  • Genetics

Background:

  • Simultaneous RNA-DNA Fluorescence In Situ Hybridization (FISH) is crucial for studying long non-coding RNAs (lncRNAs) and their genomic loci.
  • Traditional combined FISH methods often fail due to RNA signal degradation during DNA denaturation steps.

Purpose of the Study:

  • To develop a robust and simplified protocol for simultaneous RNA-DNA FISH.
  • To overcome the limitations of existing methods in preserving RNA integrity.

Main Methods:

  • Utilized amino-labeled nucleic acid probes for the RNA FISH component.
  • Integrated this with DNA FISH protocols, optimizing conditions to prevent RNA damage.

Main Results:

  • Successfully demonstrated a robust protocol for simultaneous RNA-DNA FISH.
  • The method effectively preserves fragile RNA signals during harsh DNA denaturation.

Conclusions:

  • The developed RNA-DNA FISH protocol is a significant improvement for studying RNA-DNA interactions.
  • Enables reliable detection and quantification of single RNA molecules alongside DNA in cellular environments.