Abstract
A Simple and Robust Enzymatic Fragmentation Method for NGS Library Construction Bin Li (1), James Anderson (1), Ewa Patrycja Malc (2), Piotr Mieczkowski (2), Lin Pham (1) 1 Tecan Genomics, Redwood City, CA 2 High Throughput Sequencing Facility, Department of Genetics, University of North Carolina, Chapel Hill, USA Next Generation Sequencing methods have co-evolved with sequencers to work optimally within a fairly narrow band of DNA fragment lengths. The Illumina systems require fragmenting DNA to a few hundred base pairs. Physical shearing methods, such as acoustic shearing and sonication, are the most common DNA shearing methods. Unfortunately, these methods are laborious, have increased risk of sample cross contamination, are difficult to automate and the sheared DNA requires repair for adaptor ligation and utility as a template for DNA polymerases. Earlier efforts at employing an enzymatic approach have i) required titration of enzyme to DNA ii) demonstrated cut site bias iii) resulted in a broad fragment length distribution, iv) OR all of the above. Here we present a simple and robust enzymatic method to generate random DNA fragments with compatible fragment ends for adaptor ligation. Tecan's DNA enzymatic fragmentation method is a simple add and incubate workflow that takes about 30 minutes, does not require optimization across a wide range of input amounts, provides consistent results without GC bias and is fully automatable. We show reproducible library construction on a Tecan automation workstation with consistent fragment size distribution, and sequencing results of DNA libraries with high complexity and low coverage bias. This robust and tunable DNA fragmentation method allows you to further simplify your DNA-Seq NGS library preparation workflow.