Title:

Temperature Manipulation of Small-Scale Growth Cultures for Increased pDNA Yield

The objective of this study is to investigate the capabilities of Pioreactors© – a class of small-scale bioreactors – for the optimization of experimental protocols to produce pDNA for use in our larger scale benchtop bioreactor. Pioreactors© offer a wide range of functionalities adjacent to that of a benchtop bioreactor including optical density measured via infrared light, mixing and agitation, real-time growth rate monitoring, and peristaltic pump integration. The purpose of this investigation is to develop an optimized inoculation and growth protocol that will be applied to our 7 L bioreactor. The protocol will utilize pUC plasmids, taking advantage of their ability to respond to a temperature change to increase pDNA production. Our intent is to capitalize on this characteristic to produce larger amounts of pDNA than our current protocol. The determination of an adequate protocol on a smaller scale allows for refinement before scaling to the larger bioreactor, saving on both time and cost. Through experimentation, protocol refinement, and innovative strategies, this project aims to optimize growth conditions to maximize pDNA yield for use in our larger benchtop bioreactor.

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