Efficient Production of Single-Stranded Phage DNA as Scaffolds for DNA Origami

Benjamin Kick, Florian Praetorius, Hendrik Dietz, Dirk Weuster-Botz

Research output: Contribution to journalArticlepeer-review

90 Scopus citations

Abstract

Scaffolded DNA origami enables the fabrication of a variety of complex nanostructures that promise utility in diverse fields of application, ranging from biosensing over advanced therapeutics to metamaterials. The broad applicability of DNA origami as a material beyond the level of proof-of-concept studies critically depends, among other factors, on the availability of large amounts of pure single-stranded scaffold DNA. Here, we present a method for the efficient production of M13 bacteriophage-derived genomic DNA using high-cell-density fermentation of Escherichia coli in stirred-tank bioreactors. We achieve phage titers of up to 1.6 × 1014 plaque-forming units per mL. Downstream processing yields up to 410 mg of high-quality single-stranded DNA per one liter reaction volume, thus upgrading DNA origami-based nanotechnology from the milligram to the gram scale.

Original languageEnglish
Pages (from-to)4672-4676
Number of pages5
JournalNano Letters
Volume15
Issue number7
DOIs
StatePublished - 8 Jul 2015

Keywords

  • DNA origami
  • bacteriophage M13
  • high-cell-density fermentation
  • self-assembly
  • single-stranded DNA

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