DNA-DISK: automated end-to-end data storage via enzymatic single-nucleotide DNA synthesis and sequencing on digital microfluidics

DNA is emerging as a promising alternative to conventional data storage media. However, current methods for DNA-based information storage face significant challenges, including labor-intensive processes, susceptibility to errors, and resource-intensive requirements. To make this technology more practical and accessible, we report a platform named DNA-DISK that seamlessly integrates DNA synthesis and sequencing on digital microfluidics (DMF) for data storage. The engineered Terminal deoxynucleotidyl Transferase-mediated single-nucleotide DNA synthesis coupled with bio-capping strategy is utilized, offering an eco-friendly, low-redundancy and high-fidelity approach for data writing. Pyrosequencing is employed for in-situ and accurate data reading. A DNA encapsulation solution using thermo-responsive agarose is developed for in-situ solidification, not only eliminating data clutter but also preventing DNA degradation. With DNA-DISK, the entire process of synthesis, storage, and sequencing is efficiently carried out on DMF, enabling automated liquid manipulation. As a proof of concept, DNA-DISK successfully stored and retrieved three data files, demonstrating higher storage capacity and lower write-to-read latency with superior automation and parallelization compared to other end-to-end platforms.