A Fast Solution to NGS Library Prep with Low Nanogram DNA Input

• Published in: Journal of biomolecular techniques 2013-05, Vol.24 (Suppl), p.S44-S44
• Main Authors: Liu, Pingfang, Lohman, Gregory J.S., Cantor, Eric, Langhorst, Bradley W., Yigit, Erbay, Apone, Lynne M., Munafo, Daniela B., Stewart, Fiona J., Evans, Thomas C., Nichols, Nicole, Dimalanta, Eileen T., Davis, Theodore B., Sumner, Christine
• Format: Article
• Language: English
• Subjects: Poster Session Abstracts
• ISSN: 1524-0215; 1943-4731

Next Generation Sequencing (NGS) has significantly impacted human genetics, enabling a comprehensive characterization of the human genome as well as a better understanding of many genomic abnormalities. By delivering massive DNA sequences at unprecedented speed and cost, NGS promises to make personalized medicine a reality in the foreseeable future. To date, library construction with clinical samples has been a challenge, primarily due to the limited quantities of sample DNA available. Our objective here was to overcome this challenge by developing NEBNext® Ultra DNA Library Prep Kit, a fast library preparation method. Specifically, we streamlined the workflow utilizing novel NEBNext reagents and adaptors, including a new DNA polymerase that has been optimized to minimize GC bias. As a result of this work, we have developed a simple method for library construction from an amount of DNA as low as 5 ng, which can be used for both intact and fragmented DNA. Moreover, the workflow is compatible with multiple NGS platforms.