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Related Experiment Videos

Solid phase synthesis of oligonucleotides on a crosslinked polyacrylmorpholide support

K E Norris, F Norris, K Brunfeldt

    Nucleic Acids Symposium Series
    |January 1, 1980
    PubMed
    Summary

    Researchers developed a novel polymer support for oligonucleotide synthesis using phosphodiester and phosphotriester methods. This new support facilitates efficient synthesis of DNA sequences, including a decanucleotide and an octanucleotide.

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    Area of Science:

    • Organic Chemistry
    • Biochemistry
    • Polymer Science

    Background:

    • Solid-phase synthesis is crucial for oligonucleotide production.
    • Existing polymer supports have limitations in efficiency and applicability for different chemical strategies.

    Purpose of the Study:

    • To develop and characterize a novel polymer support for oligonucleotide synthesis.
    • To demonstrate the utility of this support in both phosphodiester and phosphotriester synthesis approaches.
    • To synthesize specific DNA sequences using the developed methodology.

    Main Methods:

    • Modification of crosslinked polyacrylmorpholide beads with piperazine to create a reactive polymer support.
    • Anchoring of protected nucleotides to the polymer support via amide bond formation.

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  • Utilizing phosphodiester and phosphotriester methodologies for oligonucleotide chain elongation.
  • Devising a simplified deprotection and coupling cycle for 3' to 5' chain extension.

    • Main Results:

    • Successfully prepared a piperazine-functionalized polymer support.
    • Synthesized a self-complementary decanucleotide (d(T-C-G-G-A-T-C-C-G-A)) using the phosphodiester approach.
    • Synthesized an octanucleotide (d(T-T-T-T-T-T-T-T)) and a hexanucleotide (d(G-C-C-C-A-T)) using the phosphotriester approach.
    • Demonstrated a simple and effective deprotection and coupling cycle for chain extension.

    Conclusions:

    • The developed piperazine-functionalized polymer support is effective for both phosphodiester and phosphotriester oligonucleotide synthesis.
    • The methodology allows for the efficient synthesis of defined DNA sequences.
    • The simplified chain extension cycle enhances the practicality of solid-phase oligonucleotide synthesis.