Novel Fragmentation Method for Automated Next Generation Sequencing Exome Library Preparation

Abstract

The goal of this project was to characterize, automate, and evaluate a novel enzymatic fragmentation method for the creation of next generation sequencing libraries. Genetic sequencing through the use of next generation techniques has the potential to significantly enhance research and improve therapeutic outcomes through personalized medicine. The complexity in processing samples is one of the barriers that contributes to the high temporal and monetary cost of sequencing genetic material. New England BioLabs has recently developed the NEBnext UltraII FS system for the processing of whole genomic DNA into fragmented libraries through the use of enzymatic cleavage. This novel fragmentation technique was shown to generate libraries of comparable quality to ultrasonic shearing techniques when compared to the Agilent SureSelectXT preparation. Additionally, the new method required less sequencing depth over previous restriction enzyme based fragmentation methods to generate the same coverage metrics. The NEBnext UltraII FS system was proven to have stable results when automated allowing for rapid scalability as well as reduced cost and processing time. Ultimately, NEBnext UltraII FS is a viable candidate as the next evolution in NGS library preparation.