Simple Automated NGS Library Construction Using Optimized NEBNext(R) Reagents and a Caliper Sciclone

While next generation sequencing technologies are continually evolving to increase the data output, sequence-ready library preparation significantly lags behind in scale. The multi-step scheme of library construction and gel-based size selection limits the number of samples that can be processed manually without introducing handling errors. Moreover, processing multiple samples is extremely time consuming. Our objective here was to address these issues by developing an automated library construction process for NGS platforms. Specifically, we optimized a library construction workflow utilizing NEBNextâ reagents in conjunction with the Sciclone NGS liquid handling workstation. In addition, specific reagent configuration designs were tested for ease-of-use. Key considerations in the design of the reagent kits included the elimination of manual pipetting steps in setting up the instrument, reagent storage compatibility, the premixing of components for the various enzymatic steps and the reduction of reagent dead-volume. As a result of this work, we have developed a cost-effective automated process that is scalable from 8-96 samples with minimal hands on time.