2023 AIChE Annual Meeting

(304g) Separation and Extraction of DNA from DNA-Protein Mixtures Using a Microfluidic Device

Authors

Jiayi Wang - Presenter, University of Florida
Jason E. Butler, University of Florida
A.J.C Ladd, University of Florida
A microfluidic process utilizing a combination of flows and electricfields is used to separate long strands of DNA from proteins. Theseparation relies on DNA migrating to the bounding walls of amicrofluidic channel and then upstream against the strong pressuredriven flow due to an opposing electrophoretic velocity [1,2]. As aresult, DNA is trapped within the microfluidic channel, but proteins,which do not migrate to the bounding walls, are swept out of the channelby the convective flow. In this work, we separate mixtures of DNA(lambda phage DNA) and proteins such as bovine serum albumin (BSA).Measurements of in-situ fluorescence confirm the separation, as donepreviously [3]. Additionally, the DNA is recovered from themicrofluidic chip and analyzed. Extraction efficiency and yields arereported as a function of flow and field strengths using UV-visspectrophotometry and quantitative fluorometry. The length distributionof the recovered DNA is also reported.[1] Mert Arca, Anthony J.C. Ladd, and Jason E. Butler,“Electro-hydrodynamic concentration of genomic length DNA,” Soft Matter12, 6975-6984, 2016.[2] Ryan J. Montes, Anthony J.C. Ladd, and Jason E. Butler, “Transversemigration and microfluidic concentration of DNA using Newtonianbuffers,” Biomicrofluidics 13, 044104, 2019.[3] Benjamin E. Valley, Anne D. Crowell, Jason E. Butler, and AnthonyJ.C. Ladd, “Electrohydrodynamic extraction of DNA from mixtures of DNAand bovine serum albumin,” Analyst 145, 5532-5538, 2020.