UNIT 13.10 Gram-Scale Chemical Synthesis of 2′-Deoxynucleoside-5′-O-Triphosphates
Published Online: 1 JUN 2012
Copyright © 2012 by John Wiley & Sons, Inc.
Lab Protocol Title
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. 49:13.10.1–13.10.12.
- 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.
- chemical synthesis;
- one-pot synthesis;
- proton sponge;