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Related Concept Videos

RACE - Rapid Amplification of cDNA Ends02:35

RACE - Rapid Amplification of cDNA Ends

Rapid Amplification of cDNA Ends, or RACE, is one of the most effective methods to obtain a full-length cDNA from an mRNA sequence between a known internal region to the unknown sequence at the 5’ or 3’ end. The unknown region is cloned in the cDNA by a gene-specific primer that binds the known end, and a hybrid primer that attaches a predefined anchor sequence to the unknown end of the cDNA. The sequence in between is amplified by PCR with an anchor primer and a gene-specific primer.
Since the...
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The first human genome sequencing project cost $2.7 billion and was declared complete in 2003, after 15 years of international cooperation and collaboration between several research teams and funding agencies. Today, with the advent of next-generation sequencing technologies, the cost and time of sequencing a human genome have dropped over 100 fold.
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RNA-seq03:21

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Related Experiment Video

Updated: Jun 3, 2026

HOX Loci Focused CRISPR/sgRNA Library Screening Identifying Critical CTCF Boundaries
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Subtractive hybridization of cDNA libraries.

G S Kelner1, R A Maki

  • 1Neurocrine Biosciences, San Diego, CA.

Methods in Molecular Medicine
|March 8, 2011
PubMed
Summary

Subtractive hybridization enriches for complementary DNAs (cDNAs) that show differential gene expression between cell populations. This method is effective for identifying upregulated genes in activated cells compared to unactivated cells.

Area of Science:

  • Molecular Biology
  • Genomics
  • Biotechnology

Background:

  • Gene expression analysis is crucial for understanding cellular processes.
  • Identifying differentially expressed genes aids in discovering molecular mechanisms of cell activation.
  • Subtractive hybridization is a technique used to compare gene expression between different biological samples.

Purpose of the Study:

  • To describe the application and effectiveness of subtractive hybridization for identifying differentially expressed genes.
  • To highlight the suitability of subtractive hybridization for analyzing gene transcription changes in activated versus unactivated cell populations.

Main Methods:

  • Subtractive hybridization was employed to enrich for abundant complementary DNAs (cDNAs) that exhibit differences between two cell populations.

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  • The technique involves comparing gene transcription profiles of activated and unactivated cellular states.

    • Main Results:

    • Subtractive hybridization successfully enriches for cDNAs that are differentially expressed between cell populations.
    • The method is effective for analyzing gene expression in both homogenous and heterogeneous cell systems, including tissues.
    • The success of the technique correlates with the degree of differential gene upregulation in the activated state.

    Conclusions:

    • Subtractive hybridization is a valuable tool for isolating and identifying genes with altered expression levels.
    • The technique is particularly useful for studying gene responses to activation stimuli in various biological contexts.
    • The efficiency of subtractive hybridization is dependent on the magnitude of transcriptional differences between the compared samples.