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

Isolation of rare transcripts by representational difference analysis

M J O'Neill1, A H Sinclair

  • 1Department of Paediatrics and Centre for Hormone Research, The University of Melbourne, Royal Children's Hospital, Melbourne, Victoria 3052, Australia. oneill@cryptic.rch.unimelb.edu.au

Nucleic Acids Research
|July 1, 1997
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

Tissue magnetic susceptibility mapping as a marker of tau pathology in Alzheimer's disease.

NeuroImage·2017
Same author

A duplication in a patient with 46,XX ovo-testicular disorder of sex development refines the SOX9 testis-specific regulatory region to 24 kb.

Clinical genetics·2017
Same author

Application of neurite orientation dispersion and density imaging (NODDI) to a tau pathology model of Alzheimer's disease.

NeuroImage·2015
Same author

In vivo imaging of tau pathology using multi-parametric quantitative MRI.

NeuroImage·2015
Same author

DMRT1 is required for Müllerian duct formation in the chicken embryo.

Developmental biology·2015
Same author

Gene silencing of TNF-alpha in a murine model of acute colitis using a modified cyclodextrin delivery system.

Journal of controlled release : official journal of the Controlled Release Society·2013
Same journal

Correction to 'scSuperAnnotator: A platform for benchmarking comparison and visualizing automated cellular annotation methods for scRNA-seq data'.

Nucleic acids research·2026
Same journal

Correction to 'Differentiable partition function calculation for RNA'.

Nucleic acids research·2026
Same journal

Deployment of non-canonical splicing in tunicate genomes is mediated by divergent U2AF function and changing m6A modification in U1 and U6 snRNA.

Nucleic acids research·2026
Same journal

Bacillus subtilis DnaB forms multiple protein-protein interactions essential for DNA replication initiation.

Nucleic acids research·2026
Same journal

Multiple forms of protein-protein and DNA binding are exhibited by BrxC from the BREX phage restriction system.

Nucleic acids research·2026
Same journal

Biosynthesis of glycosylated 5-hydroxycytosine in the DNA of diverse viruses.

Nucleic acids research·2026
See all related articles

Representational difference analysis (RDA) can identify gene expression differences but often yields false positives. This study minimizes false positives and increases sensitivity in cDNA RDA, enabling rare transcript isolation from minimal mRNA.

Area of Science:

  • Molecular Biology
  • Genomics
  • Gene Expression Analysis

Background:

  • Representational difference analysis (RDA) is a technique used for genome difference cloning.
  • RDA has been adapted for identifying differentially expressed genes.
  • PCR-based differential screening methods, including RDA, are susceptible to false positives.

Purpose of the Study:

  • To identify a major source of false positives in cDNA RDA.
  • To introduce modifications to minimize false positives in cDNA RDA.
  • To increase the sensitivity of RDA for isolating rare differential transcripts.

Main Methods:

  • Utilized representational difference analysis (RDA) on cDNA.
  • Identified sources of false positives specific to cDNA RDA.

Related Experiment Videos

  • Implemented modifications to the RDA protocol.

    • Main Results:

    • A major source of false positives in cDNA RDA was identified.
    • Modifications were introduced to significantly minimize false positives.
    • The improved method demonstrated increased sensitivity, allowing isolation of rare transcripts from nanogram mRNA amounts.

    Conclusions:

    • The study successfully identified and addressed a key source of false positives in cDNA RDA.
    • The refined RDA technique enhances sensitivity and reliability for differential gene expression analysis.
    • This advancement facilitates the isolation of rare transcripts, crucial for understanding complex biological processes.