Self-assembly cloning: A rapid construction method for recombinant molecules from multiple fragments

Akira Matsumoto, Taichi Q. Itoh

Research output: Contribution to journalArticle

Abstract

Enzyme-free cloning (EFC) can rapidly produce an in-frame fusion gene with multiple fragments. To practically apply EFC, we investigated the extent and sequence of complementary staggered overhangs necessary to direct self-assembly of multiple fragments as well as a size limitation of the constructed DNA molecule. Six-base pair overhangs with 50% GC content were sufficient to direct self-assembly. A functional plasmid that exceeded 10 kb, which includes an in-frame fusion domain, was efficiently constructed from four PCR fragments in one step by our improved method.

Original languageEnglish
Pages (from-to)xxi
JournalBioTechniques
Volume62
Issue number1
DOIs
Publication statusPublished - Jan 2017

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Biochemistry, Genetics and Molecular Biology(all)

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