UNIT 3.16 A Universal and Recyclable Solid Support for Oligonucleotide Synthesis
Published Online: 1 SEP 2007
Copyright © 2007 by John Wiley and Sons, Inc.
Lab Protocol Title
Current Protocols in Nucleic Acid Chemistry
How to Cite
Morvan, F., Meyer, A. and Vasseur, J.-J. 2007. A Universal and Recyclable Solid Support for Oligonucleotide Synthesis. Current Protocols in Nucleic Acid Chemistry. 30:3.16:3.16.1–3.16.19.
- Published Online: 1 SEP 2007
- Published Print: SEP 2007
This unit provides a modified phosphoramidite method to synthesize oligodeoxyribonucleotides onto a universal and reusable hydroxyl solid support thanks to the use of deoxyribonucleoside tert-butyl and cyanoethyl phosphoramidites. The nucleoside tert-butyl phosphoramidite allows the introduction of an H-phosphonate diester linkage using the phosphoramidite method. After elongation, the H-phosphonate diester linker is cleaved by transesterification under mild basic conditions to yield an oligonucleotide with free 3′- and 5′-hydroxyls and the starting solid support. Thus, the solid support is easily recycled and used for a subsequent synthesis. In addition, a nucleoside tert-butyl phosphoramidite could be introduced inside the oligonucleotide chain during the elongation to yield a second H-phosphonate diester linkage. After elongation, the two H-phosphonate diester linkages are cleaved, producing two oligonucleotides with free 3′- and 5′-hydroxyls. Curr. Protoc. Nucleic Acid Chem. 30:3.16.1-3.16.19. © 2007 by John Wiley & Sons, Inc.
- automatized synthesis;
- tandem synthesis