Rapid, low-input, low-bias construction of shotgun fragment libraries by high-density in vitro transposition

• Main Authors: Adey, Andrew, Morrison, Hilary G, Asan, Xun, Xu, Kitzman, Jacob O, Turner, Emily H, Stackhouse, Bethany, MacKenzie, Alexandra P, Caruccio, Nicholas C, Zhang, Xiuqing, Shendure, Jay
• Format: Journal Article
• Language: English
• Published: BioMed Central Ltd 08.12.2010

© The Authors, 2010. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Genome Biology 11 (2010): R119, doi:10.1186/gb-2010-11-12-r119. This work wassupported in part by grants from the National Institutes of Health/NationalHeart Lung and Blood Institute (R01 HL094976 to JS), the National Institutesof Health/National Human Genome Research Institute (R21 HG004749 to JS),the National Institutes of Health/National Institute of Allergy and InfectiousDisease Northwest Regional Center of Excellence for Biodefense andEmerging Infectious Diseases at the University of Washington(3U54AI05714), the Ministry of Science and Technology of China, 863program (2006AA02A301), and an NSF Graduate Research Fellowship (toJOK). We characterize and extend a highly efficient method for constructing shotgun fragment libraries in which transposase catalyzes in vitro DNA fragmentation and adaptor incorporation simultaneously. We apply this method to sequencing a human genome and find that coverage biases are comparable to those of conventional protocols. We also extend its capabilities by developing protocols for sub-nanogram library construction, exome capture from 50 ng of input DNA, PCR-free and colony PCR library construction, and 96-plex sample indexing.