19. Second-Generation Sequencing Library Preparation: In Vitro Tagmentation via Transposome Insertion

  1. Dr. Matthias Harbers2,3 and
  2. Prof. Dr. Günter Kahl4,5,6
  1. Fraz Syed

Published Online: 23 JAN 2012

DOI: 10.1002/9783527644582.ch19

Tag-Based Next Generation Sequencing

Tag-Based Next Generation Sequencing

How to Cite

Syed, F. (2011) Second-Generation Sequencing Library Preparation: In Vitro Tagmentation via Transposome Insertion, in Tag-Based Next Generation Sequencing (eds M. Harbers and G. Kahl), Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim, Germany. doi: 10.1002/9783527644582.ch19

Editor Information

  1. 2

    4-2-6 Nishihara, Kashiwa-Shi, Chiba 277-0885, Japan

  2. 3

    DNAFORM Inc., Leading Venture Plaza 2, 75-1 Ono-cho, Tsurumi-ku, Yokohama, Kanagawa 230-0046, Japan

  3. 4

    Mohrmühlgasse 3, 63500 Seligenstadt, Germany

  4. 5

    University of Frankfurt am Main Biocenter, Max-von-Lauestraße 9, 60439 Frankfurt am Main, Germany

  5. 6

    Frankfurt Biotechnology Innovation Center (FIZ), GenXPro Ltd, Altenhöferallee 3, 60438 Frankfurt am Main, Germany

Author Information

  1. Epicentre Biotechnologies, 726 Post Road, Madison, WI 53713, USA

Publication History

  1. Published Online: 23 JAN 2012
  2. Published Print: 14 DEC 2011

ISBN Information

Print ISBN: 9783527328192

Online ISBN: 9783527644582

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Keywords:

  • second-generation sequencing;
  • library preparation;
  • in vitro tagmentation;
  • transposome insertion;
  • methods

Summary

The advent of next-generation sequencing (NGS) has made it possible to analyze the genome at previously unattainable depth. Numerous companies have developed well-established platforms for massively parallel deep sequencing. Regardless of the instrument, one of the bottlenecks for NGS is the amount of time and resources required for template and library preparation. Current methods generally consist of distinct steps that involve significant sample loss and hands-on time: DNA fragmentation, end-polishing, and adapter ligation. In vitro transposition with Nextera Transposomes simultaneously fragments and covalently tags the target DNA, combining these three distinct steps into a single reaction. Platform-specific adapters can be added, and the sample can be enriched and barcoded using limited-cycle polymerase chain reaction to prepare ditagged DNA fragment libraries. Nextera technology offers a streamlined, efficient, and high-throughput method for generating barcoded libraries compatible with multiple NGS platforms.