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Related Experiment Videos

Direct selection of cDNAs using whole chromosomes

S Rouquier1, B J Trask, S Taviaux

  • 1Centre de Recherche de Biochimie Macromoléculaire, CNRS UPR 9008, Montpellier, France.

Nucleic Acids Research
|November 11, 1995
PubMed
Summary

Researchers developed a new method to select complementary DNAs (cDNAs) using whole chromosomes. This technique enables rapid identification of expressed sequences from specific genomic regions, useful for chromosome painting.

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Area of Science:

  • Genomics
  • Molecular Biology
  • Biotechnology

Background:

  • Identifying specific expressed sequences within complex genomes is crucial for understanding gene function and regulation.
  • Traditional methods for isolating chromosome-specific expressed sequences can be laborious and time-consuming.

Purpose of the Study:

  • To develop a novel method for the direct selection of complementary DNAs (cDNAs) using whole chromosomes as targets.
  • To demonstrate the utility of this method for chromosome painting and rapid isolation of expressed sequences from specific genomic regions.

Main Methods:

  • Synthesis of double-strand cDNAs from human fetal brain polyadenylated messenger RNAs (mRNAs).
  • Amplification of flow-sorted chromosomes 17 and 19 using degenerate oligonucleotide primed polymerase chain reaction (DOP-PCR).

Related Experiment Videos

  • Capture of double-strand cDNAs using an improved magnetic bead protocol with amplified chromosomes as targets.

    • Main Results:

    • Selected cDNA populations specifically painted chromosomes 17 and 19 in fluorescence in situ hybridization (FISH) experiments.
    • Demonstrated the feasibility of chromosome painting using cDNA probes.
    • Confirmed the method's ability to rapidly select expressed sequences encoded by specific genomic portions.

    Conclusions:

    • The developed method allows for direct selection of chromosome-specific cDNAs.
    • This approach facilitates rapid identification of expressed sequences from any genomic region.
    • The technique is a valuable tool for chromosome painting and functional genomics research.