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

Academic Article

Abstract

  • DNA serves as a model system in polymer physics due to its ability to be obtained as a uniform polymer with controllable topology and nonequilibrium 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 producing 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. This solution of uniform supercoiled and relaxed circular plasmid DNA, is roughly 69 times greater than the overlap concentration. The utility of this method is demonstrated by performing bulk rheology measurements at sample volumes up to 1 mL on DNA of different lengths, topologies, and concentrations. 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 12 decades of frequency. Ultimately, these results can provide important insights into the dynamics of ring polymers and the nature of highly condensed DNA dynamics.
  • Authors

  • Paiva, Wynter A
  • Alakwe, Somkene D
  • Marfai, Juexin
  • Jennison-Henderson, Madigan V
  • Achong, Rachel A
  • Duche, Tinotenda
  • Weeks, April A
  • Robertson-Anderson, Rae M
  • Oldenhuis, Nate
  • Status

    Publication Date

  • August 2024
  • Published In

  • Advanced Materials  Journal
  • Keywords

  • Bioreactors
  • DNA, Circular
  • Plasmids
  • Rheology
  • bioreactors
  • circular polymers
  • plasmid DNA
  • rheology
  • unimolecular polymers
  • Digital Object Identifier (doi)

    Pubmed Id

  • 38935929
  • Start Page

  • e2405490
  • Volume

  • 36
  • Issue

  • 35