Abstract

High-throughput DNA de novo synthesis has drawn extensive attention from academia and bioindustry due to its fundamental importance in applications such as gene synthesis, drug discovery, disease diagnosis, and digital data storage. After developing for more than two decades, microchip-based DNA synthesis by inkjet printing has become one of the mainstream high-throughput DNA synthesis technologies. This technology formulates different phosphoramidite monomers into ink, then uses a multi-nozzle printing system to distribute the ink drops onto preset reactive microsites with a specific order. Therefore, the nucleotide synthetic reactions can be restricted to independent microsites with the throughput as high as several millions. Benefiting from the advantages of inkjet printing in high precision, high efficiency, and customization, this DNA technology demonstrates an appealing prospect in high-throughput and low-cost DNA synthesis. In this review, we comprehensively introduce the fundamental concepts and procedures of the inkjet printing-based DNA synthesis technology, and then summarize its essential components including the inkjet printer, the ink reagent, and the high-throughput microchip. Finally, the potential opportunities and challenges of DNA synthesis through inkjet printing are prospected. This review aims to draw more consideration to and involvement in developing high-throughput, low-cost, high-fidelity, long-strand, and efficient DNA synthesis technology.

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