Abstract
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.
Highlights
Next-generation sequencing (NGS) has significantly impacted human genetics, enabling a comprehensive characterization of human genome as well as better understanding of many genomic abnormalities
We have developed a fast library preparation method using novel NEBNext reagents and adaptors, including a new DNA polymerase that has been optimized to minimize GC bias
The workflow is compatible with multiple NGS platforms
Summary
Next-generation sequencing (NGS) has significantly impacted human genetics, enabling a comprehensive characterization of human genome as well as better understanding of many genomic abnormalities. A fast solution to NGS library preparation with low nanogram DNA input Pingfang Liu*, Gregory JS Lohman, Eric Cantor, Bradley W Langhorst, Erbay Yigit, Lynne M Apone, Daniela B Munafo, Christine Sumner, Fiona J Stewart, Thomas C Evans Jr, Nicole M Nichols, Eileen T Dimalanta, Theodore B Davis From Beyond the Genome 2012 Boston, MA, USA.
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