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

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.

SUBMITTER: Adey A

PROVIDER: S-EPMC3046479 | biostudies-literature | 2010

REPOSITORIES: biostudies-literature

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

Adey Andrew A Morrison Hilary G HG Asan Xun Xu X Kitzman Jacob O JO Turner Emily H EH Stackhouse Bethany B MacKenzie Alexandra P AP Caruccio Nicholas C NC Zhang Xiuqing X Shendure Jay J

Genome biology 20101208 12

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 constr ...[more]

PMID: 21143862

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

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