From Bioreactor to Bulk Rheology: Achieving Scalable Production of Highly Concentrated Circular DNA.

DNA serves as a model system in polymer physics due to its ability to be obtained as a uniform polymer with controllable topology and non-equilibrium behavior. Currently, a major obstacle in the widespread adoption of DNA is obtaining it on a scale and cost basis that accommodates bulk rheology and high-throughput screening. To address this, recent advancements in bioreactor-based plasmid DNA production is coupled with anion exchange chromatography to produce a unified approach to generating gram-scale quantities of monodisperse DNA. With this method, 1.1 grams of DNA is obtained per batch to generate solutions with concentrations up to 116 mg mL -1 of uniform supercoiled and relaxed circular plasmid DNA, which is roughly 69 times greater than the overlap concentration. The utility of this method is demonstrated by performing bulk rheology measurements on DNA of different length, topologies, and concentrations at sample volumes up to 1 mL. The measured elastic moduli are orders of magnitude larger than those previously reported for DNA and allowed for the construction of a time-concentration superposition curve that spans twelve decades of frequency. Ultimately, these results could provide important insights into the dynamics of ring polymers and the nature of highly condensed DNA dynamics. This article is protected by copyright. All rights reserved.