The process of oligonucleotide synthesis
The specific solid phase synthesis process of oligonucleotides consists of four cycles: detritylation, coupling, oxidation and capping, and one cycle generates a new nucleotide.
1.Detritylation
The first reaction, detritylation, removes a 5’-dimethoxytrityl protecting group from the support-bound nucleoside.
2.Coupling
The second reaction involves coupling the appropriate phosphoramidite monomer (A, C, G, or T) mixed with an activator.
3.Oxidation/thiolation
The newly-formed phosphite triester internucleotide bond is then converted to the corresponding phosphorothioate or phosphodiester with a thiolating or oxidation reagent respectively.
4.Capping
The final step in the synthesis cycle is capping the unreacted 5’-hydroxyl groups.
Applications
• Oligonucleotide drugs
• DNA sequencing primers and hybridization probes
• Synthesis of antisense nucleic acids
• Synthesis of RNA
Technical Features
STS can be used to synthesize the scale from 50 μmol to 9 mmol.
Truesynt oligonucleotide synthesizers offer:
• Flexibility in synthesis methods, chemistry, and scale
-phosphodiester and/or phosphorothioate DNA, RNA oligonucleotides
• Good synthesis of economy
- low amidite consumption
- generates significantly less waste
- re-circulation of monomer solutions
• High throughput
- cycle time for a 1 µmol synthesis is under 5 min
• Simple method scouting and scale-up
- synthesis of up to seven oligonucleotides sequentially
-simple method scale-up without additional method optimization
- STS chromatography workstation transparent to scale
• On-line monitoring for process analytical technology (PAT)
- conductivity, UV-vis, and pressure
