Preparation of next-generation sequencing libraries using Nextera™ technology: simultaneous DNA fragmentation and adaptor tagging by in vitro transposition

Methods Mol Biol. 2011;733:241-55. doi: 10.1007/978-1-61779-089-8_17.

Abstract

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.

MeSH terms

  • Base Sequence
  • DNA / chemistry*
  • DNA / genetics*
  • DNA / metabolism
  • DNA Transposable Elements / genetics*
  • Gene Library*
  • Genome / genetics
  • Sequence Analysis, DNA / methods*

Substances

  • DNA Transposable Elements
  • DNA