Accurate whole human genome sequencing using reversible terminator chemistry

被引：2388

作者：

Bentley, David R. ^{[1
]}

Balasubramanian, Shankar ^{[2
]}

Swerdlow, Harold P. ^{[1
]}

Smith, Geoffrey P. ^{[1
]}

Milton, John ^{[1
]}

Brown, Clive G. ^{[1
]}

Hall, Kevin P. ^{[1
]}

Evers, Dirk J. ^{[1
]}

Barnes, Colin L. ^{[1
,2
]}

Bignell, Helen R. ^{[1
]}

Boutell, Jonathan M. ^{[1
]}

Bryant, Jason ^{[1
]}

Carter, Richard J. ^{[1
]}

Cheetham, R. Keira ^{[1
]}

Cox, Anthony J. ^{[1
]}

Ellis, Darren J. ^{[1
]}

Flatbush, Michael R. ^{[3
]}

Gormley, Niall A. ^{[1
]}

Humphray, Sean J. ^{[1
]}

Irving, Leslie J. ^{[1
]}

Karbelashvili, Mirian S. ^{[3
]}

Kirk, Scott M. ^{[3
]}

Li, Heng ^{[4
]}

Liu, Xiaohai ^{[1
,2
]}

Maisinger, Klaus S. ^{[1
]}

Murray, Lisa J. ^{[1
]}

Obradovic, Bojan ^{[1
]}

Ost, Tobias ^{[1
]}

Parkinson, Michael L. ^{[1
]}

Pratt, Mark R. ^{[3
]}

Rasolonjatovo, Isabelle M. J. ^{[1
]}

Reed, Mark T. ^{[3
]}

Rigatti, Roberto ^{[1
]}

Rodighiero, Chiara ^{[1
]}

Ross, Mark T. ^{[1
]}

Sabot, Andrea ^{[1
]}

Sankar, Subramanian V. ^{[3
]}

Scally, Aylwyn ^{[4
]}

Schroth, Gary P. ^{[3
]}

Smith, Mark E. ^{[1
]}

Smith, Vincent P. ^{[1
]}

Spiridou, Anastassia ^{[1
]}

Torrance, Peta E. ^{[1
]}

Tzonev, Svilen S. ^{[3
]}

Vermaas, Eric H. ^{[3
]}

Walter, Klaudia ^{[4
]}

Wu, Xiaolin ^{[1
]}

Zhang, Lu ^{[3
]}

Alam, Mohammed D. ^{[3
]}

Anastasi, Carole ^{[1
]}

机构：

[1] Illumina Cambridge Ltd, Saffron Walden CB10 1XL, Essex, England

[2] Univ Cambridge, Dept Chem, Univ Chem Lab, Cambridge CB2 1EW, England

[3] Illumina Hayward, Hayward, CA 94343 USA

[4] Wellcome Trust Sanger Inst, Cambridge CB10 1SA, England

[5] Manteia Predict Med SA, CH-1267 Coinsins, Switzerland

[6] Illumina Inc, Corp Headquarters, San Diego, CA 92121 USA

[7] NHGRI, NIH, Bethesda, MD 20892 USA

来源：

NATURE | 2008年 / 456卷 / 7218期

基金：

美国国家卫生研究院; 英国惠康基金; 英国生物技术与生命科学研究理事会; 英国医学研究理事会;

关键词：

D O I：

10.1038/nature07517

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

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. 303 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.

引用

页码：53 / 59

页数：7

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