Synthesis of threose nucleic acid (TNA) triphosphates and oligonucleotides by polymerase-mediated primer extension

Su Zhang, Hanyang Yu, John C. Chaput

Research output: Contribution to journalArticlepeer-review

4 Scopus citations

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 enzymemediated polymerization of a DNA primer-template complex. Starting from suitably protected threofuranosyl nucleosides, TNA triphosphates are synthesized in a singlepot 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 9°N DNA polymerase bearing the A485L mutation.

Original languageEnglish (US)
Article number4.54
JournalCurrent Protocols in Nucleic Acid Chemistry
Issue numberSUPPL.52
DOIs
StatePublished - Mar 1 2013

Keywords

  • Alternative nucleic acids
  • Oligonucleotide
  • Polymerase-mediated primer extension
  • Threose nucleic acid (TNA)
  • Triphosphates

ASJC Scopus subject areas

  • Biochemistry
  • Organic Chemistry

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