UNIT 1.8 Synthesis, Characterization, and Application of Substituted Pyrazolopyrimidine Nucleosides
Published Online: 1 FEB 2003
Copyright © 2003 by John Wiley and Sons, Inc.
Lab Protocol Title
Current Protocols in Nucleic Acid Chemistry
How to Cite
Dempcy, R. O., Podyminogin, M. A. and Reed, M. W. 2003. Synthesis, Characterization, and Application of Substituted Pyrazolopyrimidine Nucleosides. Current Protocols in Nucleic Acid Chemistry. 11:1.8:1.8.1–1.8.15.
- Published Online: 1 FEB 2003
- Published Print: DEC 2002
This unit describes, in detail, the preparation of 3-aminopropyl-substituted pyrazolo[3,4-d]pyrimidine analogs of the purines deoxyadenosine (dA) and deoxyguanosine (dG). Phosphoramidite reagents of these so-called aminopropyl-PPA and -PPG nucleosides (AP-PPA and AP-PPG, respectively) allow introduction of amino linkers into internal positions of synthetic DNA strands. Synthesis of suitably protected AP-PPA and AP-PPG phosphoramidites are described. The stepwise alkynylation, hydrogenation, selective protection, and phosphoramidite synthesis is similar for both the PPA and PPG analogs. To demonstrate the application of these reagents, a protocol is given in which a simple DNA strand is synthesized and conjugated to a lipophilic activated ester (dabcyl-SE) to form a stable amide linkage. Utility of this chemistry for preparing internally modified DNA conjugates is discussed.