Terminator-free template-independent enzymatic DNA synthesis for digital information storage

Nat Commun. 2019 Jun 3;10(1):2383. doi: 10.1038/s41467-019-10258-1.


DNA is an emerging medium for digital data and its adoption can be accelerated by synthesis processes specialized for storage applications. Here, we describe a de novo enzymatic synthesis strategy designed for data storage which harnesses the template-independent polymerase terminal deoxynucleotidyl transferase (TdT) in kinetically controlled conditions. Information is stored in transitions between non-identical nucleotides of DNA strands. To produce strands representing user-defined content, nucleotide substrates are added iteratively, yielding short homopolymeric extensions whose lengths are controlled by apyrase-mediated substrate degradation. With this scheme, we synthesize DNA strands carrying 144 bits, including addressing, and demonstrate retrieval with streaming nanopore sequencing. We further devise a digital codec to reduce requirements for synthesis accuracy and sequencing coverage, and experimentally show robust data retrieval from imperfectly synthesized strands. This work provides distributive enzymatic synthesis and information-theoretic approaches to advance digital information storage in DNA.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Apyrase / metabolism*
  • DNA / chemical synthesis*
  • DNA Nucleotidylexotransferase / metabolism*
  • Information Storage and Retrieval / methods*
  • Nanopores
  • Sequence Analysis, DNA


  • DNA
  • DNA Nucleotidylexotransferase
  • Apyrase