title: Efficient Production of Single-Stranded Phage DNA as Scaffolds for DNA Origami.
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.
authors.affiliation: †Institute of Biochemical Engineering, Technische Universität München, Boltzmannstr. 15, 85748 Garching, Germany.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Kick
authors.affiliation: ‡Physik Department, Walter Schottky Institute, Technische Universität München, Am Coulombwall 4a, 85748 Garching, Germany.
authors.forename: Florian
authors.initials: F
authors.lastname: Praetorius
authors.affiliation: ‡Physik Department, Walter Schottky Institute, Technische Universität München, Am Coulombwall 4a, 85748 Garching, Germany.
authors.forename: Hendrik
authors.initials: H
authors.lastname: Dietz
authors.affiliation: †Institute of Biochemical Engineering, Technische Universität München, Boltzmannstr. 15, 85748 Garching, Germany.
authors.forename: Dirk
authors.initials: D
authors.lastname: Weuster-Botz
mesh_terms: D017104:Bacteriophage M13 / Q000235:genetics*; D019149:Bioreactors; D004277:DNA, Single-Stranded / Q000235:genetics*; D004279:DNA, Viral / Q000235:genetics*; D004926:Escherichia coli / Q000166:cytology / Q000235:genetics* / Q000378:metabolism*; D005285:Fermentation; D007218:Industrial Microbiology / Q000379:methods; D036103:Nanotechnology
chemical_list: D004277:DNA, Single-Stranded; D004279:DNA, Viral
keywords: DNA origami; bacteriophage M13; high-cell-density fermentation; self-assembly; single-stranded DNA