Multiple Site-Directed Mutagenesis Via Simple Cloning by Prolonged Overlap Extension | Zendy

Rasmus Hejlesen | Zendy; ErnstMartin Füchtbauer | Zendy

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Multiple Site-Directed Mutagenesis Via Simple Cloning by Prolonged Overlap Extension

Author(s) -

Rasmus Hejlesen,

ErnstMartin Füchtbauer

Publication year - 2020

Publication title -

biotechniques

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.617

H-Index - 131

eISSN - 1940-9818

pISSN - 0736-6205

DOI - 10.2144/btn-2019-0104

Subject(s) - extension (predicate logic) , mutagenesis , cloning (programming) , site directed mutagenesis , genetics , simple (philosophy) , computational biology , biology , computer science , mutation , gene , programming language , mutant , philosophy , epistemology

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.

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