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Related Experiment Video

Updated: Jul 9, 2026

Protocol for the Solid-phase Synthesis of Oligomers of RNA Containing a 2'-O-thiophenylmethyl Modification and Characterization via Circular Dichroism
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Published on: July 28, 2017

A novel catechol-based universal support for oligonucleotide synthesis.

Keith M Anderson1, Laurent Jaquinod, Michael A Jensen

  • 1Stanford Genome Technology Center, Stanford University, Palo Alto, California 94304, USA. keith.anderson@stanford.edu

The Journal of Organic Chemistry
|November 30, 2007
PubMed
Summary

A new universal support simplifies deoxyribonucleic and ribonucleic acid synthesis. This novel support enables mild cleavage and deprotection, producing purified oligonucleotides ready for biochemical assays and polymerase chain reaction (PCR) applications.

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Area of Science:

  • Oligonucleotide Synthesis
  • Biochemistry
  • Molecular Biology

Background:

  • Existing universal supports for nucleic acid synthesis often require harsh cleavage and deprotection conditions.
  • Non-volatile additives used in some methods necessitate purification steps, increasing complexity and cost.
  • The development of efficient and versatile supports is crucial for advancing molecular biology techniques.

Purpose of the Study:

  • To develop a novel universal support for both deoxyribonucleic acid (DNA) and ribonucleic acid (RNA) synthesis.
  • To improve upon existing universal supports by enabling cleavage and deprotection under mild conditions.
  • To eliminate the need for post-synthesis purification of oligonucleotides.

Main Methods:

  • Synthesis of a novel universal support from 1,4-dimethoxycatechol.
  • Characterization of oligonucleotide cleavage and deprotection kinetics using reverse-phase high-performance liquid chromatography (HPLC) and electrospray mass spectrometry.
  • Evaluation of synthesized oligonucleotide performance in polymerase chain reaction (PCR) assays.
  • Development of a method for sintering the support into filter plugs for commercial synthesizers.

Main Results:

  • The novel universal support (UL1) allows for rapid 3'-dephosphorylation under mild conditions (e.g., <1 hour in ammonium hydroxide/methylamine at 80°C).
  • Oligonucleotides synthesized using the support demonstrated full biological compatibility, performing identically to control primers in PCR assays.
  • The support plugs facilitate the synthesis of high-quality oligonucleotides up to 120 nucleotides in length with purity comparable to commercial supports.
  • Reagent consumption was reduced by approximately 50% compared to non-universal commercial supports.

Conclusions:

  • The developed universal support offers a significant advancement in oligonucleotide synthesis due to its mild cleavage and deprotection capabilities.
  • The elimination of purification steps and reduced reagent consumption enhance the efficiency and cost-effectiveness of oligonucleotide synthesis.
  • The versatility of the support for various oligonucleotide modifications and its compatibility with commercial synthesizers are expected to broaden its application in molecular biology.