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

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

Research output: Contribution to journalArticle

222 Citations (Scopus)

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.

Original languageEnglish (US)
Article numberR119
JournalGenome Biology
Volume11
Issue number12
DOIs
StatePublished - Dec 8 2010
Externally publishedYes

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transposition (genetics)
Firearms
Libraries
Exome
Transposases
Polymerase Chain Reaction
Human Genome
DNA Fragmentation
DNA
DNA fragmentation
fragmentation
genome
methodology
In Vitro Techniques
library
sampling
protocol
method

ASJC Scopus subject areas

  • Genetics
  • Cell Biology
  • Ecology, Evolution, Behavior and Systematics

Cite this

Rapid, low-input, low-bias construction of shotgun fragment libraries by high-density in vitro transposition. / Adey, Andrew; Morrison, Hilary G.; Asan; Xun, Xu; Kitzman, Jacob O.; Turner, Emily H.; Stackhouse, Bethany; MacKenzie, Alexandra P.; Caruccio, Nicholas C.; Zhang, Xiuqing; Shendure, Jay.

In: Genome Biology, Vol. 11, No. 12, R119, 08.12.2010.

Research output: Contribution to journalArticle

Adey, A, Morrison, HG, Asan, Xun, X, Kitzman, JO, Turner, EH, Stackhouse, B, MacKenzie, AP, Caruccio, NC, Zhang, X & Shendure, J 2010, 'Rapid, low-input, low-bias construction of shotgun fragment libraries by high-density in vitro transposition', Genome Biology, vol. 11, no. 12, R119. https://doi.org/10.1186/gb-2010-11-12-r119
Adey, Andrew ; Morrison, Hilary G. ; Asan ; Xun, Xu ; Kitzman, Jacob O. ; Turner, Emily H. ; Stackhouse, Bethany ; MacKenzie, Alexandra P. ; Caruccio, Nicholas C. ; Zhang, Xiuqing ; Shendure, Jay. / Rapid, low-input, low-bias construction of shotgun fragment libraries by high-density in vitro transposition. In: Genome Biology. 2010 ; Vol. 11, No. 12.
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