This PFI:AIR Technology Translation project focuses on nucleic acid fractionation to fill the need for adequate sample preparation for the rapidly increasing field of next generation genome sequencing. The microfluidic nucleic acid sorter is important because genome sequencing is fundamental for deciphering biological pathways leading to malfunction and disease. All sequencing technologies require the fractionation of nucleic acids tailored to the specific sequencing approach. This project will result in a proof-of-concept design for a microfluidic nucleic acid sorter. This microfluidic nucleic acid sorter has the following unique features: a dielectrophoretic process fractionating nucleic acids by size and conformation, realization on a microfluidic platform and operation under continuous flow. These features provide the advantages of nucleic acid fractionation in tunable size ranges, cost effectiveness, minute sample amounts and applicability to both DNA and RNA when compared to the leading competing shearing technology and bead purification technology in this market space.
This project addresses previously little explored technology gaps in next generation sequencing as it translates from research discovery toward commercial application, comprised of the development of a versatile device applicable to a wide range of nucleic acid lengths, conformations and nucleic acid types and, the micro-preparative approach realized through microfluidic integration. Specifically, the project aims in the development of nucleic acid array and constriction fractionators based on insulator-based dielectrophoresis suitable for fragment lengths between 100 bp and ~20 kbp and in volumes and quantities applicable for next generation sequencing. The design of the dielectrophoretic fractionation process will be optimized with the aid of numerical simulation tools and improved device fabrication technologies. A systematic study of size fractionation capabilities of the proof-of-principle devices for DNA and RNA will be performed and optimized fractions will be tested on existing next generation sequencing platforms. The versatility of the approach will allow the application for existing but also future next generation sequencing platforms and their requirements in nucleic acid sample preparation.
In addition, personnel involved in this project, undergraduate students and post docs, will receive innovation and technology translation experiences through active involvement in the research components of this project, the proof-of-principle development and the collaboration with an industry partner. Mentoring activities further fosters the participation of women in the research aspects of this project.
