UNIT 13.10 Gram-Scale Chemical Synthesis of 2′-Deoxynucleoside-5′-O-Triphosphates

  1. Anilkumar R. Kore,
  2. Muthian Shanmugasundaram,
  3. Annamalai Senthilvelan,
  4. Balasubramanian Srinivasan

Published Online: 1 JUN 2012

DOI: 10.1002/0471142700.nc1310s49

Current Protocols in Nucleic Acid Chemistry

Current Protocols in Nucleic Acid Chemistry

How to Cite

Kore, A. R., Shanmugasundaram, M., Senthilvelan, A. and Srinivasan, B. 2012. Gram-Scale Chemical Synthesis of 2′-Deoxynucleoside-5′-O-Triphosphates. Current Protocols in Nucleic Acid Chemistry. 49:13.10:13.10.1–13.10.12.

Author Information

  1. Bioorganic Chemistry Division, Life Technologies Corporation, Austin, Texas

Publication History

  1. Published Online: 1 JUN 2012


A simple, straightforward, reliable, and efficient method for the chemical synthesis of sodium salt of 2′-deoxynucleoside-5′-O-triphosphates (dNTPs), starting from the corresponding nucleoside, is described. This improved “one-pot, three-step” synthetic strategy involves the monophosphorylation of nucleoside, followed by reaction with tributylammonium pyrophosphate and hydrolysis of the resulting cyclic intermediate to provide the corresponding dNTP in good yields (65% to 70%). It is noteworthy that the protocol holds good for both the purine deoxynucleotides, such as 2′-deoxyguanosine-5′-O-triphosphate (dGTP) and 2′-deoxyadenosine-5′-O-triphosphate (dATP), and pyrimidine deoxynucleotides, such as 2′-deoxycytidine-5′-O-triphosphate (dCTP), thymidine-5′-O-triphosphate (TTP), and 2′-deoxyuridine-5′-O-triphosphate (dUTP). Curr. Protoc. Nucleic Acid Chem. 49:13.10.1-13.10.12. © 2012 by John Wiley & Sons, Inc.


  • 2′-deoxyribonucleotides;
  • chemical synthesis;
  • one-pot synthesis;
  • phosphorylation;
  • proton sponge;
  • triphosphate;
  • dNTP