High Quality Library Construction and Reliable Quantitation with NEBNext Reagents

The field of next generation sequencing (NGS) has matured significantly over the past few years. As the quantity of data generated per NGS run increases and the time required per run decreases, the ability to quickly produce large numbers of high quality libraries inexpensively is becoming increasingly important. Moreover, maximizing data output & minimizing instrument run failure are imperative. In this poster, we present data showing: 1) the improvements we have made in library preparation with the development of the NEBNext Ultra II Library Prep kit; and 2) the development of the NEBNext Library Quant Kit for Illumina, a simple and robust method for quantitation of Illumina libraries. In the first part, we show that libraries made with the Ultra II kit have low DNA input requirements; additionally, we significantly improved library yields and reduced sequence bias. Moreover, the workflow is simple and streamlined, greatly reducing the time required to produce high quality DNA libraries and the possibility of errors. In the second part, we demonstrate the effectiveness of the NEBNext Library Quant Kit for a broad range of library types and sizes as well as advantages offered by qPCR quantitation for obtaining optimal cluster density and user-to-user consistency. The NEBNext Quant Kit offers an efficient and cost-effective qPCR library quantitation workflow for users looking to optimize both sequencing yield and throughput.