Related Experiment Videos
A short, novel, and cheaper procedure for oligonucleotide synthesis using automated solid phase synthesizer
Quanlai Song1, Zhiwei Wang, Yogesh S Sanghvi
1Isis Pharmaceuticals, Inc., Carlsbad, California 92008, USA.
Nucleosides, Nucleotides & Nucleic Acids
|October 21, 2003
Summary
Dimethylthiuram disulfide (DTD) offers efficient thiolation for oligonucleotide synthesis. This method streamlines the process, reducing solvent use and synthesis time for large-scale production.
Area of Science:
- Oligonucleotide Synthesis
- Chemical Reagents
- Biochemistry
Background:
- Automated oligonucleotide synthesis relies on phosphoramidite chemistry.
- Efficient thiolation reagents are crucial for producing phosphorothioate oligonucleotides.
- Current methods can be time-consuming and solvent-intensive.
Purpose of the Study:
- To introduce Dimethylthiuram disulfide (DTD) as an efficient thiolation reagent.
- To develop a streamlined synthesis protocol for phosphorothioate oligonucleotides.
- To demonstrate the feasibility of large-scale synthesis using DTD.
Main Methods:
- Utilized Dimethylthiuram disulfide (DTD) in phosphoramidite chemistry for oligonucleotide synthesis.
- Combined thiolation and capping steps by mixing DTD with capping solution B.
- Performed large-scale synthesis of phosphorothioate oligonucleotides at the 1 mmol scale.
Main Results:
- Achieved simultaneous thiolation and capping, reducing solvent consumption by 20%.
- Compressed the standard four-step synthesis cycle into three steps.
- Demonstrated high yield and purity in large-scale (1 mmol) synthesis of phosphorothioate oligonucleotides.
Conclusions:
- Dimethylthiuram disulfide (DTD) is an effective reagent for automated oligonucleotide synthesis.
- The developed method offers significant advantages in efficiency and resource conservation.
- This approach enables robust large-scale production of phosphorothioate oligonucleotides.