A Novel Instrument and Consumables for Quality Control in Next Generation Sequencing

ABSTRACT Sample preparation for next-generation sequencing (NGS), also known as library construction, is a laborious and lengthy process consisting of numerous complex biochemical operations that require several quality control (QC) checks to ensure optimal success. Gel electrophoresis utilizing automated instrumentation remains the standard QC technique for nearly all NGS applications and library workflows, whereby several nano-grams of DNA at numerous library QC checkpoints is separated by size, visualized via fluorescence and compared to known standards (DNA ladders) for analysis. With the rapid advancement of NGS technologies and library preparation chemistries, gel electrophoresis has fallen behind the critical market needs of cost reduction, faster analysis time, lower sample input (sensitivity), reduced complexity of kits and cold chain reagents, sample structural information, and sizing resolution of large DNA. Evizia has developed PRECYSE, a biomolecule desktop imaging system based on High-Speed Atomic Force Microscopy (HSAFM), through combining expertise across physics, chemistry, genomics and software analytics. A user replaceable AFM probe scans an atomically flat consumable surface containing up to 96 separate DNA samples to generate a topographical image down to single molecule nanometer resolution. Onboard software instantaneously analyses complex biomolecule mixtures to generate vastly improved QC metric specificity and sensitivity. Specific capabilities include sizing of DNA from 100 bp to 250 kb+ in minutes, consolidation of QC to a single inexpensive consumable chip without cold chain reagents, <25 pico-grams of input DNA, and the unique capability of directly visualizing structural information; bound proteins, DNA secondary structure, or library contaminants. Initial customer collaborations showed numerous structural DNA phenomena undetectable by electrophoresis that could directly impact sequencing throughput and read quality. Evizia aims to refine consumable production, further optimize key instrument components for further cost reduction and continue expanding customer NGS collaborations. PRECYSE will redefine how NGS QC is performed today for all customer archetypes. In exceeding NGS QC market demands Evizia hopes to contribute to the rapid expansion of NIH funded NGS programs looking to advance our understanding of the human genome and develop personalized medicine.