Integrated system for preparation of NGS libraries from crude biological samples

DESCRIPTION (provided by applicant): The overall goal for the proposed SBIR program is to develop an instrument system that can accept whole blood as an input sample and produce a genomic DNA library suitable for next- generation DNA sequencing (NGS). Allsteps, from DNA extraction to final library construction, take place by a single automated process, without any user intervention. Because of the fully automated process, the proposed system would greatly lower the cost and labor of NGS sequencing, and accelerate movement of NGS technology into clinical diagnostic settings. Another advantage of the instrument is that it should be scalable from 100's of ml of blood down to the single cell level. Application of NGS technology to small samples is an increasingly important goal in cancer research and diagnostics. The proposed system utilizes a novel approach to microfluidic sample preparation. A single continuous-flow technology is used to manipulate and separate cells, subcellular organelles, and large genomicDNA molecules (that exhibit particle-like properties). The use of a common technology for all sample prep steps provides several benefits: 1) Multiple sequential processing steps can be accomplished in a single operation, on a single consumable device, thereby simplifying system design. 2) In multi-step processes, integration of reaction steps and post-reaction cleanup steps enables seamless, potentially zero-loss transfer of sample between processing steps. 3) Sample purification is accomplished on thebasis of particle size alone. Differential adsorption to solid phases is not used, and sample loss due to incomplete elution is voided. 4) It is straightforward to automate the lengthy, complex sample preparation protocols that are typical of most NGSplatforms, making it easier and more cost effective to move NGS technology into high throughput applications like clinical trials research and diagnostic testing. PUBLIC HEALTH RELEVANCE: The overall goal for the proposed SBIR program is to develop an instrument system that can accept whole blood as an input sample and produce a genomic DNA library suitable for next- generation DNA sequencing (NGS). All steps, from DNA extraction to final library construction, take place by a single automated process, without any user intervention. Because of the fully automated process, the proposed system would greatly lower the cost and labor of NGS sequencing, and accelerate movement of NGS technology into clinical diagnostic settings.