Single restriction enzyme-assisted megaprimer polymerase chain reaction to fuse two DNA sequences on separate cloning vectors

Nobuhisa Umeki, Taro Q.P. Uyeda

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

We describe a simple and versatile method to fuse two DNA sequences on separate cloning vectors in a single polymerase chain reaction (PCR). The method, termed restriction enzyme-assisted megaprimer PCR (REM-PCR), requires that the two cloning vectors share a common sequence and that the DNA sequences to be fused are cloned in the same orientation with respect to the common sequence. Fusion of the two sequences is achieved by mutual priming at the common sequence between two DNA fragments that were generated by restriction enzyme and linearly amplified by repetitive priming in the PCR reaction mixture.

Original languageEnglish
Pages (from-to)309-311
Number of pages3
JournalAnalytical Biochemistry
Volume414
Issue number2
DOIs
Publication statusPublished - 2011 Jul 15
Externally publishedYes

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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