Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

Ribonuclease protection.

J L Thorvaldsen1, M S Bartolomei

  • 1Howard Hughes Medical Institute and Department of Cell and Developmental Biology, University of Pennsylvania School of Medicine, Philadelphia, PA, USA.

Methods in Molecular Biology (Clifton, N.J.)
|July 5, 2003
PubMed
Summary
This summary is machine-generated.

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

TET1 non-catalytic activity shapes the chromatin landscape that directs de novo methylation establishment in the male germline.

bioRxiv : the preprint server for biology·2025
Same author

Transgenerational effects of maternal bisphenol A exposure on offspring metabolic health - Erratum.

Journal of developmental origins of health and disease·2019
Same author

Transgenerational effects of maternal bisphenol: a exposure on offspring metabolic health.

Journal of developmental origins of health and disease·2018
Same author

Transgenerational inheritance of chronic adolescent stress: Effects of stress response and the amygdala transcriptome.

Genes, brain, and behavior·2018
Same author

Establishment and maintenance of H19 imprinting in the germline and preimplantation embryo.

Cytogenetic and genome research·2006
Same author

Genomic imprinting: antagonistic mechanisms in the germ line and early embryo.

Cold Spring Harbor symposia on quantitative biology·2005
Same journal

Tracking Synthetic Adhesins on Bacterial Surfaces with Immunofluorescence Microscopy.

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

Post-Selection Methods for Analyzing mRNA Display Selections and Optimization of Hits.

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

High-Performance Computing in Tandem Mass Spectrometry (MS/MS) Peptide Identification.

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

Engineering and Adapting Disulfide-Containing Proteins to Enable Intracellular Functionality.

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

AI-Driven Protein Research: From Prediction to Design.

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

Methods for the In Vitro Selection of Protein and Peptide Libraries Using mRNA Display.

Methods in molecular biology (Clifton, N.J.)·2026
See all related articles

The ribonuclease protection assay (RPA) offers sensitive detection of total cellular RNA, outperforming Northern analysis and directly analyzing RNA without reverse transcription. This method is quantitative and can detect allele-specific gene expression.

Area of Science:

  • Molecular Biology
  • Genetics

Background:

  • The ribonuclease protection assay (RPA) is a sensitive method for analyzing total cellular RNA.
  • It offers advantages over Northern analysis in terms of sensitivity and tolerance to RNA degradation.

Purpose of the Study:

  • To detail the methodology and advantages of the ribonuclease protection assay (RPA) for RNA analysis.
  • To highlight its utility in studying imprinted genes and allele-specific expression.

Main Methods:

  • Generating a specific antisense riboprobe.
  • Hybridizing the probe to total RNA.
  • RNase digestion of unprotected RNA, followed by isolation and analysis of protected RNA on a denaturing gel.

Main Results:

  • RPA demonstrates high sensitivity, detecting as little as 0.1 pg of target RNA.

Related Experiment Videos

  • The assay is more tolerant of partially degraded RNA compared to other methods.
  • RPA allows direct analysis of RNA, bypassing the need for reverse transcription.
  • The method is quantitative when the probe is in excess.

    • Conclusions:

    • RPA is a sensitive, quantitative, and versatile technique for total cellular RNA analysis.
    • Its ability to detect allele-specific expression makes it particularly valuable for studying imprinted genes.