Research progress in high-throughput DNA synthesis and its applications.

In recent years, the development of high-throughput DNA synthesis technology has significantly advanced research in genomics and synthetic biology. Traditional DNA synthesis methods, such as first-generation DNA synthesizer and PCR-based approaches, have demonstrated excellent performance in many aspects. However, they exhibit notable limitations in synthesis of long-chain DNA and large-scale parallel synthesis. Second-generation high-throughput DNA synthesis technologies, including photolithographic, inkjet, electrochemical, and thermally controlled synthesis techniques based on microarray chips, have shown remarkable advantages in improving synthesis efficiency, reducing costs, and increasing throughput. However, these methods rely on chemical principles, making it challenging to overcome issues related to short sequence length and environmental pollution. This has led to the emergence of third-generation enzymatic synthesis technologies, which offer distinct advantages in environmental sustainability and long-chain DNA synthesis, demonstrating great application potential. This review defines microarray-based synthesis as the boundary for high-throughput synthesis, categorizing previous methods as traditional synthesis technologies. It systematically elaborates on mainstream high-throughput synthesis technologies, analyzing and comparing their advantages and limitations. Furthermore, it explores their applications in life sciences, medicine, and other fields. Finally, potential technological advancements and application expansions are discussed, providing insights into the future development directions and challenges of high-throughput DNA synthesis technology, with the aim of offering valuable references for related research.