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Multiple site-directed mutagenesis via simple cloning by prolonged overlap extension

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We describe the application of simple cloning by prolonged overlap extension for multiple site-directed mutagenesis in the same plasmid. We show that it is possible to use this technique with very short PCR templates. The technique is ideally suited for the generation of longer donor DNA sequences for CRISPR/Cas9-mediated homologous repair.

Original languageEnglish
Pages (from-to)345-348
Number of pages4
Publication statusPublished - Jun 2020

    Research areas

  • donor template generation, gene editing, multiple site-directed mutagenesis, prolonged overlap extension, seamless cloning, seamless plasmid assembly

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ID: 190965274