Unit

UNIT 4.54 Synthesis of Threose Nucleic Acid (TNA) Triphosphates and Oligonucleotides by Polymerase-Mediated Primer Extension

  1. Su Zhang,
  2. Hanyang Yu,
  3. John C. Chaput

Published Online: 1 MAR 2013

DOI: 10.1002/0471142700.nc0454s52

Current Protocols in Nucleic Acid Chemistry

Current Protocols in Nucleic Acid Chemistry

How to Cite

Zhang, S., Yu, H. and Chaput, J. C. 2013. Synthesis of Threose Nucleic Acid (TNA) Triphosphates and Oligonucleotides by Polymerase-Mediated Primer Extension. Current Protocols in Nucleic Acid Chemistry. 52:4.54:4.54.1–4.54.17.

Author Information

  1. Center for Evolutionary Medicine and Informatics, The Biodesign Institute, and Department of Chemistry and Biochemistry at Arizona State University, Tempe, Arizona

Publication History

  1. Published Online: 1 MAR 2013

Abstract

This unit describes the chemical synthesis of α-l-threofuranosyl nucleic acid (TNA) triphosphates for thymidine (T), guanosine (G), cytidine (C), and the diaminopurine (D) analog of adenosine and their incorporation into TNA oligonucleotides by enzyme-mediated polymerization of a DNA primer-template complex. Starting from suitably protected threofuranosyl nucleosides, TNA triphosphates are synthesized in a single-pot reaction and purified by ion-exchange and HPLC chromatography. Purified TNA triphosphates are diluted into stock solutions and used as substrates for the synthesis of TNA oligonucleotides. Oligonucleotide synthesis is accomplished using Therminator DNA polymerase, a commercial variant of the 9oN DNA polymerase bearing the A485L mutation. Curr. Protoc. Nucleic Acid Chem. 52:4.54.1-4.54.17. © 2013 by John Wiley & Sons, Inc.

Keywords:

  • alternative nucleic acids;
  • threose nucleic acid (TNA);
  • triphosphates;
  • oligonucleotide;
  • polymerase-mediated primer extension