UNIT 3.12 Nucleoside Phosphoramidites Containing Cleavable Linkers
Published Online: 1 JAN 2006
Copyright © 2005 by John Wiley and Sons, Inc.
Lab Protocol Title
Current Protocols in Nucleic Acid Chemistry
How to Cite
Pon, R. T. 2006. Nucleoside Phosphoramidites Containing Cleavable Linkers. Current Protocols in Nucleic Acid Chemistry. 23:3.12:3.12.1–3.12.24.
- Published Online: 1 JAN 2006
- Published Print: DEC 2005
Phosphoramidite reagents (linker phosphoramidites) containing a cleavable 3′-ester linkage between the nucleoside and the phosphoramidite group can be used to attach the first nucleoside to a solid-phase support. Inexpensive underivatized supports such as LCAA-CPG can then be used as universal supports for oligonucleotide synthesis. No modifications to synthesis coupling conditions and no 3′-dephosphorylation are required. Only oligonucleotides with terminal 3′-OH ends are produced. Phosphoramidites containing both a succinate and a sulfonyldiethanol linkage are particularly useful and create oligonucleotides with both a 3′-OH and 5′-phosphate. In addition, by using these reagents, one oligonucleotide sequence can be added onto the 5′-end of another (tandem synthesis) to produce a string of multiple oligonucleotides linked end-to-end. Deprotection releases the oligonucleotides from each other to yield a mixture of oligonucleotides. This approach is particularly useful for making pairs of PCR primers or both strands of a double-stranded sequence in a single operation.
- linker phosphoramidites;
- solid-phase supports;
- tandem oligonucleotide synthesis;
- 96-well plate synthesis;
- high-throughput synthesis;
- support derivatization