Abstract

New storage technologies are needed to keep up with the global demands of data generation. DNA is an ideal storage medium due to its stability, information density and ease-of-readout with advanced sequencing techniques. However, progress in writing DNA is stifled by the continued reliance on chemical synthesis methods. The enzymatic synthesis of DNA is a promising alternative, but thus far has not been well demonstrated in a parallelized manner. Here, we report a multiplexed enzymatic DNA synthesis method using maskless photolithography. Rapid uncaging of Co2+ ions by patterned UV light activates Terminal deoxynucleotidyl Transferase (TdT) for spatially-selective synthesis on an array surface. Spontaneous quenching of reactions by the diffusion of excess caging molecules confines synthesis to light patterns and controls the extension length. We show that our multiplexed synthesis method can be used to store digital data by encoding 12 unique DNA oligonucleotide sequences with video game music, which is equivalent to 84 trits or 110 bits of data.

Highlights

  • New storage technologies are needed to keep up with the global demands of data generation

  • We show that our multiplex synthesis method can be used to store digital data by encoding 12 unique DNA oligonucleotides sequences with 84 trits (110 bits) of video game music in non-identical base transitions

  • The photolithographic activation of Terminal deoxynucleotidyl Transferase (TdT) is achieved by optically controlling the ‘spatiotemporal’ concentration distribution of its metal cofactor cobalt (Co2+) near the array surface with the photocleavable caging molecule DMNP-EDTA17–19

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Summary

Introduction

New storage technologies are needed to keep up with the global demands of data generation. Incubation with optimized ratios of apyrase, TdT, and nucleotide over multiple cycles resulted in the successful synthesis of several DNA oligonucleotides comprised of short homopolymeric blocks encoding digital information within unique base transitions.

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