Preparation of Next-Generation Sequencing Libraries Using Nextera™ Technology: Simultaneous DNA Fragmentation and Adaptor Tagging by In Vitro Transposition

Part of the Methods in Molecular Biology book series (MIMB, volume 733)


DNA library preparation is a common entry point and bottleneck for next-generation sequencing. Current methods generally consist of distinct steps that often involve significant sample loss and hands-on time: DNA fragmentation, end-polishing, and adaptor-ligation. In vitro transposition with Nextera™ Transposomes simultaneously fragments and covalently tags the target DNA, thereby combining these three distinct steps into a single reaction. Platform-specific sequencing adaptors can be added, and the sample can be enriched and bar-coded using limited-cycle PCR to prepare di-tagged DNA fragment libraries. Nextera technology offers a streamlined, efficient, and high-throughput method for generating bar-coded libraries compatible with multiple next-generation sequencing platforms.

Key words

Next-generation sequencing DNA library preparation Roche/454 Illumina/Solexa Nextera 


  1. 1.
    Kahvehian, V., Quackenbush, J., and Thompson, J. F. (2008) What would you do if you could sequence everything? Nature Biotechnology. 26, 1125–1133.CrossRefGoogle Scholar
  2. 2.
    Metzker, M. L. (2010) Sequencing Technologies – the next generation. Nature Reviews – Genetics. 11, 31–  46.Google Scholar
  3. 3.
    Shendure, J. and Henlee, J. (2008) Next-generation DNA Sequencing. Nature Biotechnology. 26, 1135–1145.PubMedCrossRefGoogle Scholar
  4. 4.
    Mardis, E. (2008) Next-Generation DNA Sequencing Methods. (2008) Annual Review of Genomics and Human Genetics. 9, 387–  402.Google Scholar
  5. 5.
    Fuller, C. W., Meddendorf, L. R., Benner, S. A., Church, G. M., Harris, T., Huang, X., Jovanovich, S. B., Nelson, J. R., Schloss, J. A., Schwartz, D. C., and Vezenov, D. V. (2009) The challenges of sequencing by synthesis. Nature Biotechnology. 27, 1013  –1023.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  1. 1.Epicentre BiotechnologiesMadisonUSA

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