Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by

Same author

[Association between genetic polymorphism of tumor necrosis factor and chronic severe hepatitis B in patients].

Zhonghua yi xue za zhi·2007

Same author

In vivo translational inaccuracy in Escherichia coli: missense reporting using extremely low activity mutants of Vibrio harveyi luciferase.

Biochemistry·2007

Same author

[Construction of recombinant adenovirus vector expressing extracellular domain of TbetaR-II-RANTES fusion gene and its anti-tumor effects].

Zhonghua zhong liu za zhi [Chinese journal of oncology]·2007

Same author

[Characteristics, evolution and variation of M genes of human avian H5N1 strains in Guangdong].

Bing du xue bao = Chinese journal of virology·2007

Same author

Dynamic changes in microbial activity and community structure during biodegradation of petroleum compounds: a laboratory experiment.

Journal of environmental sciences (China)·2007

Same author

Differences in optical transport properties between human meridian and non-meridian.

The American journal of Chinese medicine·2007

Same journal

Mapping the 3D Chromosome Organization of a Biosynthetic Gene Cluster by Capture Hi-C (CHi-C).

Methods in molecular biology (Clifton, N.J.)·2026

Same journal

Mapping the 3D Chromosome Organization of Streptomyces by Hi-C.

Methods in molecular biology (Clifton, N.J.)·2026

Same journal

CUT&Tag Epigenomic Profiling of Biosynthetic Gene Clusters in Arabidopsis thaliana.

Methods in molecular biology (Clifton, N.J.)·2026

Same journal

Rhizobium rhizogenes-Mediated Hairy Root Transformation Protocol for Lotus japonicus and Other Legumes.

Methods in molecular biology (Clifton, N.J.)·2026

Same journal

Characterization of Bioactive Saponins from Sea Cucumbers.

Methods in molecular biology (Clifton, N.J.)·2026

Same journal

Methods for Functional Validation of Terpenoid Metabolic Clusters in Nicotiana benthamiana and Aspergillus oryzae.

Methods in molecular biology (Clifton, N.J.)·2026

See all related articles

Search research articles

Related Experiment Video

Updated: Jan 3, 2026

Antibody-Free Assay for RNA Methyltransferase Activity Analysis

Antibody-Free Assay for RNA Methyltransferase Activity Analysis

Published on: July 9, 2019

RNase Protection Assay.

Jianzhu Zhao¹, Jun Tang², Justin Elfman³

¹Department of Oncology, Shengjing Hospital of China Medical University, Shenyang, China.

Methods in Molecular Biology (Clifton, N.J.)

|November 16, 2019

Summary

This summary is machine-generated.

The RNase Protection Assay (RPA) validates chimeric RNAs by detecting double-stranded RNA formed with labeled probes. This method avoids reverse transcription, preventing false positives and ensuring accurate RNA analysis.

Keywords:

Chimeric RNA Denaturing polyacrylamide gel electrophoresis Double-stranded RNA RNase Radioactively labeled Single-stranded RNA

More Related Videos

Semi-quantitative Detection of RNA-dependent RNA Polymerase Activity of Human Telomerase Reverse Transcriptase Protein

Semi-quantitative Detection of RNA-dependent RNA Polymerase Activity of Human Telomerase Reverse Transcriptase Protein

Published on: June 12, 2018

Fluorescence Based Primer Extension Technique to Determine Transcriptional Starting Points and Cleavage Sites of RNases In Vivo

Fluorescence Based Primer Extension Technique to Determine Transcriptional Starting Points and Cleavage Sites of RNases In Vivo

Published on: October 31, 2014

Related Experiment Videos

Last Updated: Jan 3, 2026

Antibody-Free Assay for RNA Methyltransferase Activity Analysis

Antibody-Free Assay for RNA Methyltransferase Activity Analysis

Published on: July 9, 2019

Semi-quantitative Detection of RNA-dependent RNA Polymerase Activity of Human Telomerase Reverse Transcriptase Protein

Semi-quantitative Detection of RNA-dependent RNA Polymerase Activity of Human Telomerase Reverse Transcriptase Protein

Published on: June 12, 2018

Fluorescence Based Primer Extension Technique to Determine Transcriptional Starting Points and Cleavage Sites of RNases In Vivo

Fluorescence Based Primer Extension Technique to Determine Transcriptional Starting Points and Cleavage Sites of RNases In Vivo

Published on: October 31, 2014

Area of Science:

Molecular Biology
Biochemistry
Genetics

Background:

Single-stranded RNA is degraded by RNase, while double-stranded RNA is resistant.
Chimeric RNAs require accurate validation methods.
Reverse transcription can introduce false positives in RNA analysis.

Purpose of the Study:

To present the RNase Protection Assay (RPA) as a method for validating chimeric RNAs.
To highlight the advantage of RPA in avoiding reverse transcription-associated false positives.

Main Methods:

Generating RNA probes labeled with 32P or biotin, complementary to target chimeric RNA.
Hybridizing labeled probes to RNA samples to form double-stranded RNA hybrids.
Utilizing RNase digestion to remove undigested single-stranded RNA, leaving protected hybrids.
Identifying protected double-stranded RNA hybrids via high-resolution, denaturing polyacrylamide gel electrophoresis.

Main Results:

RNase Protection Assay (RPA) successfully validates chimeric RNAs.
The assay demonstrates resistance of double-stranded RNA to RNase digestion.
Absence of reverse transcription in RPA circumvents potential false-positive results.

Conclusions:

RNase Protection Assay (RPA) is a reliable method for chimeric RNA validation.
RPA offers an accurate alternative to methods involving reverse transcription.
The assay's principle of RNase resistance ensures specific detection of chimeric RNAs.