Multiplexed DNA Assembly from Oligo-pools
Track: Main track (up to 8 pages)
Abstract: In recent years, DNA synthesis technologies has served as a powerful tool to validate hypothesis, greatly advanced many research areas. From simpliy validating the functionality of AI-generated genetic circuitus to deepening our understanding on life's basic principles by synthesizing a genome. However, the cost of synthesizing gene-sized DNA sequences at scale remains too high, making it economically unfeasible to validate the experimental performance of ML models or generate new datasets for futher training. To address this challenge, we developed a novel method for the high-throughput assembly of gene-sized DNA sequences, starting from cost-effective chip-synthesized oligo-pools. Unlike traditional Polymerase Cycling Assembly (PCA) methods, we employed Golden Gate Assembly (GGA) to facilitate the ligation of short DNA fragments. This approach enabled us to successfully assemble high-quality DNA libraries containing up to 96 gene-sized sequences (~600 bp) in a single-pot reaction, with convenient retrieval of individual sequences. Leveraging automation to conduct numerous reactions in parallel, we can efficiently and economically synthesize gene-sized DNA sequences at scale, thereby facilitating the experimental data generation for the ML community.
Submission Number: 74
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