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

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

Author information

1
Department of Genetics, Harvard Medical School, Boston, 02115, MA, USA. hhlee@genetics.med.harvard.edu.
2
Wyss Institute for Biologically Inspired Engineering at Harvard University, Boston, 02115, MA, USA. hhlee@genetics.med.harvard.edu.
3
Department of Genetics, Harvard Medical School, Boston, 02115, MA, USA.
4
Wyss Institute for Biologically Inspired Engineering at Harvard University, Boston, 02115, MA, USA.
5
Technicolor Research & Innovation Lab, Palo Alto, 94306, CA, USA.
6
Department of Genetics, Harvard Medical School, Boston, 02115, MA, USA. gchurch@genetics.med.harvard.edu.
7
Wyss Institute for Biologically Inspired Engineering at Harvard University, Boston, 02115, MA, USA. gchurch@genetics.med.harvard.edu.

Abstract

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.

PMID:
31160595
PMCID:
PMC6546792
DOI:
10.1038/s41467-019-10258-1
[Indexed for MEDLINE]
Free PMC Article

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