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Enzyme-Catalyzed Strategies for the Efficient Synthesis of DNA Fragments

Author(s)

Ruoshui Du1,*, Zitong Zeng2, Louis Shen3

Affiliation(s)

1Millbrook School, Millbrook, NY 12545, USA 2Sendelta International Academy, No. 6, Shenzhen, China 3Beijing SMIC Private School, Beijing, China *Corresponding author

Abstract

This work explores using synthetic DNA fragment synthesis enzymes to achieve a high yield with minimal error rate and high accuracy. DNA fragment synthesis is a fundamental genetic engineering, synthetic biology, and molecular diagnostics process. However, unlike chemical methods, enzyme-based methods are advantageous because they are specific but incur costs, efficiency, and scalability problems, making them less attractive. Different enzymes, such as polymerases, ligases, and nucleases, are studied regarding the outcome of factors like enzyme concentration, temperature, and buffer conditions on DNA synthesis. Key performance metrics such as yield, error rate, and reaction time were analyzed using techniques like PCR ligation and restriction digestion, which were used to analyze experimentally. Results show that optimizing reaction conditions leads to much greater efficiency with high-fidelity polymerases, giving greater accuracy at yield cost. For example, Taq polymerase offers higher yields but more errors. Likewise, optimized conditions resulted in a complemented outcome in ligation and restriction digestion reactions. DNA fragment synthesis is proven to rely upon enzyme-catalyzed methods, where the successful evolution in enzyme technology is driving toward more reliable and economical applications in biotechnology.

Keywords

Enzyme Catalysis; DNA Synthesis; Polymerases; Ligases; Nucleases; Molecular Biology

References

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