Amplicon Shearing Using a Covaris® Focused-ultrasonicator
The advent of next generation sequencing (NGS) has transformed the utility of genomics particularly in the field of clinical diagnostics. Recent technological advances in high-throughput sequencing applications have aided in the selection of effective personalized therapies based off an individual’s genetic makeup. Despite the major advantages provided by DNA sequencing, the generation of quality sequencing data can be significantly affected by a myriad of factors including: quality of input sample, sample mass, and sample preparation methodology used. One of the most significant factors that can affect sequencing is the DNA fragmentation step. Inefficient fragmentation can result in inefficient clustering efficiency thereby producing poor sequencing readouts. In this application note, we describe the use of Covaris Adaptive Focused Acoustics® (AFA®) technology for mechanical DNA shearing of amplicons ranging in size from 2 kb – 400 bp to 400 bp – 150 bp in preparation for massively parallel high-throughput sequencing.