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

Genome Biol. 2010;11(12):R119. doi: 10.1186/gb-2010-11-12-r119. Epub 2010 Dec 8.


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.

Publication types

  • Comparative Study
  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • DNA / isolation & purification
  • DNA Fragmentation*
  • Drosophila / genetics
  • Escherichia coli / genetics
  • Exons
  • Genome, Human
  • Genome, Insect
  • Genomic Library*
  • Humans
  • Polymerase Chain Reaction / methods*
  • Sequence Analysis, DNA / methods*
  • Transposases / metabolism


  • DNA
  • Transposases