Rapid, low-input, low-bias construction of shotgun fragment libraries by high-density in vitro transposition |
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Authors: | Andrew Adey Hilary G Morrison Asan Xu Xun Jacob O Kitzman Emily H Turner Bethany Stackhouse Alexandra P MacKenzie Nicholas C Caruccio Xiuqing Zhang Jay Shendure |
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Affiliation: | Department of Genome Sciences, University of Washington, Seattle, WA 98195, USA. acadey@uw.edu |
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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. |
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