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

被引：396

作者：

Adey, Andrew ^{[2
]}

Morrison, Hilary G. ^{[3
]}

Asan ^{[1
]}

Xun, Xu ^{[1
]}

Kitzman, Jacob O. ^{[2
]}

Turner, Emily H. ^{[2
]}

Stackhouse, Bethany ^{[2
]}

MacKenzie, Alexandra P. ^{[2
]}

Caruccio, Nicholas C. ^{[4
]}

Zhang, Xiuqing ^{[1
]}

Shendure, Jay ^{[2
]}

机构：

[1] BGI Shenzhen, Shenzhen 518000, Peoples R China

[2] Univ Washington, Dept Genome Sci, Seattle, WA 98195 USA

[3] Marine Biol Lab, Woods Hole, MA 02543 USA

[4] Epictr Biotechnol, Madison, WI 53713 USA

来源：

GENOME BIOLOGY | 2010年 / 11卷 / 12期

基金：

美国国家卫生研究院;

关键词：

AMPLIFICATION-FREE; SHORT-READ; SEQUENCE; DNA; TN5; SEQ;

D O I：

10.1186/gb-2010-11-12-r119

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

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.

引用

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