Exome Sequencing Facilitated by A Fast Library Preparation Method with Low Nanogram DNA Input

Next Generation Sequencing (NGS) has significantly impacted the field of genetics, enabling a comprehensive characterization of genomes of all living species. Exome sequencing is an effective and cheaper alternative to whole genome sequencing, and its use in clinical diagnostic applications is rapidly increasing. Recent reports of the diagnostic use of exome sequencing have shown its ability in identifying disease-causing mutations for conditions such as congenital deafness and intellectual disabilities, which were formerly undiagnosable. The hope is that such sequencing will guide physicians on the proper course of treatment, determine personal response to a drug, assess risks for siblings, and possibly predict disease before it starts. A major challenge of exome sequencing-based clinical diagnosis is the limited quantities of sample DNA available. To overcome this challenge, we have developed a fast library preparation method using novel NEBNext reagents and adaptors, including a new DNA polymerase that has been optimized to minimize GC bias. This method enables exome sequencing of libraries constructed from low nanogram quantities of DNA amplified with limited PCR cycles, and can be used for both intact and fragmented DNA including FFPE and circulating plasma DNA. Moreover, the workflow is compatible with multiple NGS platforms.