Feasibility on the synthesis of trimer based DNA sequences for combinatorial DNA encoding in data скачать в хорошем качестве

Feasibility on the synthesis of trimer based DNA sequences for combinatorial DNA encoding in data

This study demonstrates the feasibility of applying combinatorial DNA encoding for DNA-based data storage. We focus on synthesizing DNA sequences using a combination of Bz-NPPOC and DMT phosphoramidites, emphasizing the incorporation of DMT phosphoramidites as trimers. Specifically, we used the trimer “TAC” and tested its integration, showing that more than half of the synthesized sequences contained one trimer compared to control sequences synthesized with only Bz-NPPOC monomers. However, as the number of consecutive trimers increased, their presence in the sequences decreased, as expected. This reduction is due to the less efficient deprotection of DMT phosphoramidites, which are deprotected using TCA, while Bz-NPPOC phosphoramidites are deprotected with UV light. This differential deprotection mechanism complicates the synthesis of sequences containing both monomers and trimers, requiring optimization to prevent incomplete deprotection and sequence loss. Additionally, depurination during TCA exposure may cleave some sequences and remove them from further processing. Despite these challenges, we employed Maskless Array Synthesis (MAS) for high-throughput synthesis of large sequence libraries. This method uses a digital micromirror device to spatially pattern light from a UV LED onto the synthesis surface, enabling precise, sequence-defined DNA oligonucleotide synthesis. MAS allows the creation of numerous new sequence combinations by incorporating a mixture of trimers, significantly increasing the data storage density. The system can produce over two million sequences in parallel on a single array. By selectively deprotecting Bz-NPPOC phosphoramidites at different spots on the array, MAS ensures that DMT trimers couple only with those sites, incorporating them accurately into the sequences. This approach is particularly effective for combinatorial DNA encoding, offering scalability and flexibility for creating large, high-density, and error-resistant DNA libraries. This work sets the stage for developing scalable and efficient DNA storage solutions by synthesizing mixtures of monomers and trimers, overcoming deprotection challenges. Presented by Hamed Sabzalipoor, University of Munich This is a presentation from the 2025 Storage and Computing with DNA Conference. • Learn More about the SNIA DNA Data Storage Alliance: https://www.snia.org/groups/snia-dna-... • SNIA Educational Library: https://snia.org/library • X:   / snia   • LinkedIn:   / snia