A Fast Solution for NGS Library Prep with Low Nanogram DNA Input, for Multiple Sample Types and Sequencing Platforms

As the quantity of data generated per next generation sequencing (NGS) run increases and the time required per run decreases, the ability to quickly produce large numbers of libraries inexpensively is becoming increasingly important.   For example, sequencing runs on the MiSeq can take as little as 8 hours.  Moreover, the increased throughput per run enables the simultaneous sequencing of multiple samples. These advances in massively parallel DNA sequencing have resulted in library construction becoming a bottleneck to sequencing.  Additionally, library construction can be challenging due to limited quantities and low quality of some samples. To overcome these challenges, we have developed, a fast, streamlined DNA library preparation method using novel reagents and adaptors, including a DNA polymerase that has been optimized to minimize GC bias. This method enables library construction starting with nanogram quantities of DNA, and can be used for both intact and fragmented DNA. This method has been evaluated with a variety of plant and animal samples, and is also compatible with multiple NGS platforms, enabling use of a single library prep kit for creation of a variety of sequencing libraries. Illumina, 454, and SOLiD 5500 libraries were successfully generated and sequenced on the corresponding platforms to produce high quality sequencing data.