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

Andrew Adey1, Hilary G Morrison2, Asan3, Xu Xun3, Jacob O Kitzman1, Emily H Turner1, Bethany Stackhouse1, Alexandra P MacKenzie1, Nicholas C Caruccio4, Xiuqing Zhang3* and Jay Shendure1*

Author Affiliations

1 Department of Genome Sciences, University of Washington, Seattle, WA 98195, USA

2 The Marine Biological Laboratory, Woods Hole, MA 02543, USA

3 BGI-Shenzhen, Shenzhen 518000, China

4 Epicentre Biotechnologies, 726 Post Road, Madison, WI 53713, USA

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Genome Biology 2010, 11:R119  doi:10.1186/gb-2010-11-12-r119

Published: 8 December 2010

Abstract

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.