Scaling down bioreactor processes to create unimolecular supercoiled, cyclic, and linear DNA for study of topology effects in soft materials.

Biomass DNA materials have recently gained traction due to declining DNA costs and their relatively underexplored applications at bulk scales. Due to the demand for nucleic acid therapeutics, bioreactor based production has tremendously increased the yield of plasmid DNA to > 1 g of purified plasmid DNA (pDNA) per liter of culture fluid. In this work we discuss reverse engineering industrial scale fermentation and purification strategies to access gram scale amounts of pDNA which will act as precursors to DNA hydrogel materials with controlled topological elements. Beyond its secondary structure, dsDNA exhibits exquisite variety in its geometric shape and topological form, but we have not yet tapped the full potential of this feature as a tool to fine-tune and control the properties of materials on a molecular scale. As these topological forms influence virtually every DNA process, vast literature precedent dedicated to understanding the role of DNA topology in life processes would serve as the ideal foundation for a unique material. We report up to 100 mg/mL solutions of unimolecular DNA.