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

High-throughput screening of suppression subtractive hybridization cDNA libraries using DNA microarray analysis.

Noëlani van den Berg1, Bridget G Crampton, Ingo Hein

  • 1University of Pretoria, Pretoria, South Africa.

Biotechniques
|November 25, 2004
PubMed
Summary

This study introduces a quantitative method to screen complementary DNA (cDNA) libraries made using suppression subtractive hybridization (SSH). This approach effectively identifies differentially expressed genes in plants like pearl millet and banana.

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

Evaluating LED parameters in a growth chamber to maintain potato yield and enhance minitubers production in greenhouse with molecular insights.

Scientific reports·2026
Same author

The oomycete effector-targeted nuclear envelope protein StKAKU4 negatively regulates plant immunity via vesicle trafficking.

Plant physiology·2026
Same author

3RAD-Guided SNP Discovery for Species Identification and Conservation of the Medicinal Southern African Tree Genus <i>Greyia</i> Hook. & Harv.

Ecology and evolution·2026
Same author

An improved Solanum verrucosum genome provides insight into potato centromeres and epigenetic regulation.

G3 (Bethesda, Md.)·2026
Same author

Phased potato genome assembly and association genetics enable delineation of the H1 resistance locus against potato cyst nematodes.

TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik·2026
Same author

Isolation and proteomic analysis of intracellular vesicles from the potato late blight pathogen Phytophthora infestans.

Scientific reports·2026

Area of Science:

  • Molecular Biology
  • Gene Expression Analysis
  • Plant Science

Background:

  • Differential gene expression is crucial for understanding biological processes.
  • Suppression subtractive hybridization (SSH) is a common technique for isolating differentially expressed transcripts.
  • Efficient screening of SSH-generated cDNA libraries is essential for accurate downstream analysis.

Purpose of the Study:

  • To develop and validate a quantitative procedure for screening cDNA libraries constructed via SSH.
  • To identify up-regulated transcripts and assess transcript abundance in SSH libraries.
  • To apply the methodology to pearl millet and banana SSH libraries.

Main Methods:

  • Quantitative screening of SSH cDNA libraries using two-color cyanin dye labeling and hybridization.

Related Experiment Videos

  • Calculation of enrichment ratios (ER1 and ER2) for each clone.
  • Graphical representation of ER1 and ER2 to identify differentially expressed clones.
  • Normalization of clones based on ER2 values to determine transcript abundance.
  • Verification of differential expression using inverse Northern blot analysis.

    • Main Results:

    • The quantitative procedure successfully identified clones likely representing up-regulated transcripts.
    • ER1 and ER2 values provided insights into transcript enrichment and abundance.
    • The method was effectively applied to pearl millet and banana SSH libraries.
    • Differential expression was confirmed for selected clones.

    Conclusions:

    • The presented quantitative screening method is effective for analyzing SSH-constructed cDNA libraries.
    • This technique facilitates the identification of differentially expressed genes in plants.
    • The approach aids in distinguishing rare from abundant transcripts within libraries.