A Low-Cost Platform for Routine High-Accuracy Sizing of Nucleic Acid Samples with 100 bp - 100 Mbp Dynamic Range

The goal of this project is to develop a microand nanofluidic DNA sizing platform with a sensitivityspeedand dynamic range that is unmatched by current sizing technologiesWe find that fluorescence burst analysis of individual DNA molecules transported through sub micrometer scale channels provides accurate and precise sizing of fragments betweenbp ands of MbpIn this singlemolecule sizing platformunlike in gel or capillary electrophoresisthe same instrumentdeviceand set of operating conditions produce optimal sizing performance across the full dynamic rangeThis capability in a low costrobust platform will be useful for many assays that require the measurement of DNA size distributions in heterogeneous solutionsGiven the ability to rapidly size DNA molecules longer thankbpthe platform is particularly valuable for the quality control of long read sequencing librariesimproving the efficiency of sequencing workflowsConsumable fluidic devices that allow the precise control of DNA transport can be fabricated using injection molding of thermoplasticsIn this projectwe will define the optimal device designs and operating conditions to maximize the accuracyprecisionsensitivityand dynamic range of this high throughput platformOnce these parameters are defined in Phasewe will enhance the stability and reproducibility of sizing runs in Phaseof the projectThis will be done through a combination of mechanical and optical engineering and the use of sizing markersSizing performance will be validated against pulsed field gel electrophoresisThe project will culminate in a demonstration of platform utility in characterizing the quality of high mass DNA before and after library preparation for long read sequencing The goal of this project is to develop a microand nanofluidic DNA sizing platform with a sensitivityspeedand dynamic range that is unmatched by current sizing technologiesCurrent technologies do not allow the sizing of DNA molecules larger thankbp with a resolution or speed optimized for the quality control of long read sequencing librariesWe have demonstrated a single molecule nanofluidic sizing platform that can accurately size DNA fragments betweenbp ands of Mbp and can improve sequencing workflows and the efficiency of genomic studies