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Related Concept Videos

FISH - Fluorescent In-situ Hybridization02:07

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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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Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
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Related Experiment Video

Updated: Jun 26, 2025

Detection of Viral RNA by Fluorescence in situ Hybridization FISH
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Published on: May 5, 2012

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RNA-FISH for HIV Transcription/Localization Analysis.

Pavitra Ramdas1, Ajit Chande2

  • 1Molecular Virology Laboratory, Department of Biological Sciences, Indian Institute of Science Education and Research, Bhopal, Madhya Pradesh, India. pavitra17@iiserb.ac.in.

Methods in Molecular Biology (Clifton, N.J.)
|May 14, 2024
PubMed
Summary

This study details a protocol for RNA fluorescence in situ hybridization (FISH) to detect human immunodeficiency virus type 1 (HIV-1) RNA transcripts in cells. The method aids in understanding viral transcription during infection.

Keywords:
HIV-1 transcriptionQuantification of viral transcriptsRNA-FISHTranscript localization

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Area of Science:

  • Molecular Biology
  • Virology
  • Cell Biology

Background:

  • RNA fluorescence in situ hybridization (FISH) is crucial for visualizing specific RNA molecules within cells.
  • FISH is applicable for observing messenger RNA (mRNA) linked to significant genes and viral transcription in infected cells.

Purpose of the Study:

  • To provide a comprehensive protocol for the detection, localization, and quantification of HIV-1 transcripts in mammalian cell lines.
  • To enhance understanding of molecular processes during HIV-1 infection, focusing on the transcription phase.

Main Methods:

  • The protocol involves reagent preparation, cellular treatments, and visualization techniques.
  • Data acquisition and subsequent analysis are integral parts of the methodology.

Main Results:

  • The developed protocol enables precise detection and localization of HIV-1 transcripts.
  • Quantitative analysis of viral RNA is achievable using this FISH method.

Conclusions:

  • This protocol offers a valuable tool for studying HIV-1 transcription dynamics.
  • Understanding viral transcription is key to comprehending the HIV-1 life cycle and developing interventions.