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

Nano Lett. 2015 Jul 8;15(7):4672-6. doi: 10.1021/acs.nanolett.5b01461. Epub 2015 Jun 3.

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 × 10(14) 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.

Keywords: DNA origami; bacteriophage M13; high-cell-density fermentation; self-assembly; single-stranded DNA.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacteriophage M13 / genetics*
  • Bioreactors
  • DNA, Single-Stranded / genetics*
  • DNA, Viral / genetics*
  • Escherichia coli / cytology
  • Escherichia coli / genetics*
  • Escherichia coli / metabolism*
  • Fermentation
  • Industrial Microbiology* / methods
  • Nanotechnology

Substances

  • DNA, Single-Stranded
  • DNA, Viral