UNIT 4.54 Synthesis of Threose Nucleic Acid (TNA) Triphosphates and Oligonucleotides by Polymerase-Mediated Primer Extension
Published Online: 1 MAR 2013
Copyright © 2013 by John Wiley & Sons, Inc.
Lab Protocol Title
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.
- Published Online: 1 MAR 2013
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.
- alternative nucleic acids;
- threose nucleic acid (TNA);
- polymerase-mediated primer extension