Researchers from Aarhus University have developed a new and efficient way to streamline the process of synthetic DNA production.
The production of synthetic oligonucleotides is critical for a range of different areas. Their availability is one of the cornerstones of biotechnology. They are also a prerequisite for several revolutionary technologies like PCR, DNA sequencing, synthetic biology and CRISPR-Cas9. Let’s not forget how critical PCR has been for the identification of SARS-CoV-2! Chemical modification of oligonucleotides has also been important in antisense oligonucleotide-based strategies which are showing promise for the treatment of several diseases. Overall, this emphasises the ongoing high demand for oligonucleotides.
Phosphoramidite chemistry for DNA synthesis has been the gold standard for synthesis of oligonucleotides. This process however relies on phosphoramidites which are unstable unless stored at -20 degrees Celsius. Current instruments used for DNA synthesis are not able to cool down the phosphoramidites and therefore some degrade after being added to the instrument.
Improving bench stability
In a recent paper, published in Nature, researchers have proposed a new method for the preparation of phosphoramidites. In this method, the phosphitylating reagent is immobilised on a solid support. This enables rapid phospitylation of nucleosides that are flushed through a solid support. This support also ensures that nucleosides are converted to phosphoramidites within a few minutes. These are then automatically flushed on to the part of the instrument involved in DNA synthesis.
This method avoids the degradation of phosphoramidites as they are being produced just before they are used. This allows for a faster and more efficient approach that the team hope could be integrated into DNA synthesisers for an automatic approach. This would omit manual synthesis and storage of phosphoramidites. The team also noted that this setup is easy-to-use and could be operated by non-chemists.
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