Accurate whole human genome sequencing using reversible terminator chemistry

作者：David Bentley, Shankar Balasubramanian, Harold Swerdlow, Geoff Smith, John Milton, Clive G. Brown, Kevin P. Hall, Dirk J. Evers, Colin L. Barnes, Helen Bignell, Jonathan M. Boutell, Jason E. Bryant, Richard J. Carter, R. Keira Cheetham, Anthony J. Cox, Darren J. Ellis, Michael R. Flatbush, Niall A. Gormley, Sean Humphray, Leslie J. Irving, Mirian Karbelashvili, Scott M. Kirk, Heng Li, Xiaohai Liu, Klaus S. Maisinger, Lisa J. Murray, Bojan Obradović, Tobias W. B. Ost, Michael L. Parkinson, Mark R. Pratt, Isabelle Rasolonjatovo, Mark T. Reed, Roberto Rigatti, Chiara Rodighiero, Mark T. Ross, Andrea Sabot, S. V. Sankar, Aylwyn Scally, Gary P. Schroth, Mark E. B. Smith, Vincent P. Smith, Anastassia Spiridou, Peta E. Torrance, Svilen Tzonev, Eric H. Vermaas, Klaudia Walter, Xiaolin Wu, Lu Zhang, Mohammed D. Alam, Carole Anastasi, Ify C. Aniebo, David M. Bailey, Iain R. Bancarz, Saibal Banerjee, Selena G. Barbour, Primo A. Baybayan, Vincent A. Benoit, Kevin F. Benson, Claire Bevis, Phillip J. Black, Asha Boodhun, Joe S. Brennan, John A. Bridgham, Rob Brown, Andrew Anand Brown, Dale H. Buermann, Abass A. Bundu, James C. Burrows, Nigel P. Carter, Néstor Castillo-Magallanes, Maria Chiara E. Catenazzi, Simon Chang, Rachel Cooley, Natasha R. Crake, Olubunmi O. Dada, Konstantinos D. Diakoumakos, Belen Dominguez‐Fernandez, David J. Earnshaw, Ugonna C. Egbujor, David W. Elmore, Sergey Etchin, Mark Ewan, Milan Fedurco, Louise J. Fraser, Karin V. Fuentes Fajardo, W. Scott Furey, David George, Kimberley J. Gietzen, Colin P. Goddard, George S. Golda, Philip A. Granieri, David E. Green, David Gustafson, Nancy Fisher Hansen, Kevin Harnish, Christian D. Haudenschild, Narinder I. Heyer, Matthew M. Hims, Johnny T. Ho, Adrian M. Horgan, Katya Hoschler, Steve Hurwitz, D. V. Ivanov, Maria Q. Johnson, Terena James, Ted Jones, Gyoung‐Dong Kang, Tzvetana H. Kerelska, Alan D. Kersey, Irina Khrebtukova, Alex Kindwall, Zoya Kingsbury, Paula I. Kokko-Gonzales, Anil Kumar, Marc Laurent, Cindy Lawley, Sarah Emerson Lee, Xavier Lee, Arnold Liao, Jennifer A. Loch, Mitch Lok, Shujun J. Luo, Radhika M. Mammen, John William. Martin, Patrick G. McCauley, Paul McNitt, Parul D. Mehta, Keith Moon, Joe W. Mullens, Taksina Newington, Zemin Ning, Bee Ling Ng, Sonia M Novo, Michael J. O’Neill, Mark Andrew Osborne, Andrew P. Osnowski, Omead Ostadan, Lambros L. Paraschos, Lea Pickering, Andrew C. Pike, Alger C. Pike, D. Chris Pinkard, Daniel P. Pliskin, Joe Podhasky, Victor J. Quijano, Come Raczy, Vicki H. Rae, Stephen R. Rawlings, Ana Chiva Rodriguez, Phyllida Roe, John H. ROGERS, M. Candelaria Rogert Bacigalupo, Nikolai S. Romanov, Anthony Romieu, Rithy K. Roth, Natalie J. Rourke, Silke T. Ruediger, Eli Rusman, Raquel Sanches-Kuiper, Martin Schenker, Josefina M. Seoane, Richard J. Shaw, Mitch K. Shiver, S. W. Short, Ning L. Sizto, Johannes P. Sluis, Melanie A. Smith, Jean-Ernest Sohna Sohna, Eric J. Spence, Kim Stevens, Neil Sutton, Lukasz Szajkowski, Carolyn Tregidgo, Gerardo Turcatti, Stephanie vandeVondele, Yuli Verhovsky, Selene M. Virk, Suzanne Wakelin, Gregory C. Walcott, Jingwen Wang, Graham J. Worsley, Juying Yan, Ling Yau, Mike Zuerlein, Jane Rogers, James C. Mullikin, Matthew E. Hurles, Nick J. McCooke, John S. West, Frank L. Oaks, Peter L. Lundberg, David A. Klenerman, Richard M. Durbin, Anthony J. Smith · 发表于：Nature · 年份：2008 · DOI：10.1038/nature07517 · 被引用次数：3808 · 研究领域：Genomic variations and chromosomal abnormalities、Genomics and Phylogenetic Studies、Chromosomal and Genetic Variations

DNA sequence information underpins genetic research, enabling discoveries of important biological or medical benefit. Sequencing projects have traditionally used long (400–800 base pair) reads, but the existence of reference sequences for the human and many other genomes makes it possible to develop new, fast approaches to re-sequencing, whereby shorter reads are compared to a reference to identify intraspecies genetic variation. Here we report an approach that generates several billion bases of accurate nucleotide sequence per experiment at low cost. Single molecules of DNA are attached to a flat surface, amplified in situ and used as templates for synthetic sequencing with fluorescent reversible terminator deoxyribonucleotides. Images of the surface are analysed to generate high-quality sequence. We demonstrate application of this approach to human genome sequencing on flow-sorted X chromosomes and then scale the approach to determine the genome sequence of a male Yoruba from Ibadan, Nigeria. We build an accurate consensus sequence from >30× average depth of paired 35-base reads. We characterize four million single-nucleotide polymorphisms and four hundred thousand structural variants, many of which were previously unknown. Our approach is effective for accurate, rapid and economical whole-genome re-sequencing and many other biomedical applications. The power of the latest massively parallel synthetic DNA sequencing technologies is demonstrated in two major collaborations t...