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Rapid gene splicing and multi-sited mutagenesis by one-step overlap extension polymerase chain reaction.

Author information

Affiliations

  • 1. Lauren Biotechnology, Nanjing 210019, China.
      Authors
    • Wei H 1
    (1 author)

Analytical Biochemistry, 04 Jul 2012, 429(1):76-78
https://doi.org/10.1016/j.ab.2012.06.027 PMID: 22771440 

Abstract 

Gene splicing and site-directed mutagenesis (SDM) are important to introduce desired sequences in target DNA. However, introducing mutations at multiple sites requires multiple steps of DNA manipulation, which is time-consuming and labor-intensive. Here, we present a rapid efficient gene splicing and multi-sited mutagenesis method that introduces mutations at two distant sites via sequential connection of DNA fragments by one-step overlap extension polymerase chain reaction (OE-PCR). This bottom-up approach for DNA engineering can be broadly used to study protein structure-function, to optimize codon use for protein expression, and to assemble genes of interest.

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Read article at publisher's site: https://doi.org/10.1016/j.ab.2012.06.027

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