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Related Concept Videos

Ribosome Profiling02:24

Ribosome Profiling

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.
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique helps...

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Related Experiment Video

Updated: Jul 7, 2026

Method for the Isolation and Identification of mRNAs, microRNAs and Protein Components of Ribonucleoprotein Complexes from Cell Extracts using RIP-Chip
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Method for the Isolation and Identification of mRNAs, microRNAs and Protein Components of Ribonucleoprotein Complexes from Cell Extracts using RIP-Chip

Published on: September 29, 2012

Ribonuclease protection assay.

M Gilman1

  • 1Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, USA.

Current Protocols in Molecular Biology
|February 12, 2008
PubMed
Summary
This summary is machine-generated.

High-activity hybridization probes are generated using bacteriophage RNA polymerase for efficient transcription of cloned sequences. This method enables sensitive detection of target mRNA through ribonuclease protection assays and gel electrophoresis analysis.

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A Rapid High-throughput Method for Mapping Ribonucleoproteins (RNPs) on Human pre-mRNA
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A Rapid High-throughput Method for Mapping Ribonucleoproteins (RNPs) on Human pre-mRNA

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Last Updated: Jul 7, 2026

Method for the Isolation and Identification of mRNAs, microRNAs and Protein Components of Ribonucleoprotein Complexes from Cell Extracts using RIP-Chip
13:34

Method for the Isolation and Identification of mRNAs, microRNAs and Protein Components of Ribonucleoprotein Complexes from Cell Extracts using RIP-Chip

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Capture and Identification of RNA-binding Proteins by Using Click Chemistry-assisted RNA-interactome Capture (CARIC) Strategy
09:36

Capture and Identification of RNA-binding Proteins by Using Click Chemistry-assisted RNA-interactome Capture (CARIC) Strategy

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A Rapid High-throughput Method for Mapping Ribonucleoproteins (RNPs) on Human pre-mRNA
13:00

A Rapid High-throughput Method for Mapping Ribonucleoproteins (RNPs) on Human pre-mRNA

Published on: December 2, 2009

Area of Science:

  • Molecular Biology
  • Biotechnology
  • Nucleic Acid Chemistry

Background:

  • Developing highly specific and active hybridization probes is crucial for accurate nucleic acid detection.
  • Traditional probe preparation methods can be time-consuming and yield probes with variable specific activity.

Purpose of the Study:

  • To develop a robust method for generating sequence-specific hybridization probes with high specific activity.
  • To enable sensitive and reliable detection of target messenger RNA (mRNA) in biological samples.

Main Methods:

  • Cloning probe sequences downstream of a bacteriophage promoter on a plasmid.
  • In vitro transcription using bacteriophage RNA polymerase for high-abundance RNA probe synthesis.
  • Purification of RNA probes via removal of DNA template and unincorporated labels or gel electrophoresis.
  • Hybridization of probe RNA to sample RNA, followed by ribonuclease treatment to degrade unbound probe.
  • Analysis of protected probe fragments by gel electrophoresis to identify target mRNA.

Main Results:

  • Efficient and high-yield transcription of cloned sequences into discrete RNA species.
  • Purified RNA probes demonstrated high specific activity.
  • The ribonuclease protection assay coupled with gel electrophoresis successfully identified target mRNA based on the size of protected probe fragments.

Conclusions:

  • This method provides a reliable approach for producing high-specific-activity RNA probes.
  • The developed technique allows for sensitive and specific detection of target mRNA.
  • This strategy has significant implications for gene expression analysis and diagnostics.