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

SSH adequacy to preimplantation mammalian development: scarce specific transcripts cloning despite irregular

L C Bui1, R D Léandri, J P Renard

  • 1UMR Biologie du Développement et de la Reproduction, INRA 78350 Jouy en Josas, France. bui.linh-chi@jouy.inra.fr

BMC Genomics
|November 10, 2005
PubMed
Summary

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

In vitro production significantly reduces metabolic differences among bovine embryos.

Metabolomics : Official journal of the Metabolomic Society·2025
Same author

Coprophagia in early life tunes expression of immune genes after weaning in rabbit ileum.

Scientific reports·2024
Same author

52 Equine embryo size does matter!

Reproduction, fertility, and development·2022
Same author

Impact of the polycarbonate strippers used in assisted reproduction techniques on embryonic development.

Human reproduction (Oxford, England)·2020
Same author

Erratum. Maternal ageing impairs mitochondrial DNA kinetics during early embryogenesis in mice.

Human reproduction (Oxford, England)·2019
Same author

Maternal ageing impairs mitochondrial DNA kinetics during early embryogenesis in mice.

Human reproduction (Oxford, England)·2019

Suppression subtractive hybridization (SSH) effectively identifies rare, specific genes in small samples like early embryos. A reporter transcript method reliably assesses SSH library quality and repeatability for gene discovery.

Area of Science:

  • Molecular Biology
  • Genomics
  • Developmental Biology

Background:

  • Suppression subtractive hybridization (SSH) is a key technique for identifying differentially expressed genes.
  • It is particularly useful for low-abundance transcripts and when starting with limited RNA, such as in early mammalian embryo development.
  • Assessing the quality and repeatability of SSH libraries is crucial for systematic gene characterization.

Purpose of the Study:

  • To develop and validate a reliable strategy for assessing SSH library quality and repeatability.
  • To investigate the performance of SSH when using small amounts of RNA, as in early embryo development.
  • To confirm the enrichment of rare, tester-specific transcripts in SSH libraries.

Main Methods:

  • Construction of four independent SSH libraries from rabbit and bovine embryos.

Related Experiment Videos

  • Large-scale screening of clones using exogenous reporter transcripts to assess SSH procedure performance.
  • Sequencing of clones to validate results for both exogenous and endogenous transcripts.

    • Main Results:

    • Normalization in SSH is most effective for rare and moderately abundant transcripts; abundant transcripts can escape this process.
    • Some abundant transcripts common to tester and driver samples may not be effectively subtracted.
    • SSH libraries were significantly enriched for very rare, tester-specific transcripts (0.0005% of mRNAs).

    Conclusions:

    • The reporter transcript follow-up method provides a simple and reliable way to evaluate SSH performance.
    • Despite occasional normalization and subtraction issues, SSH consistently enriches libraries with rare, specific transcripts.
    • SSH is a powerful tool for gene discovery in biological contexts with limited material, like early mammalian development.