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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.

OriginalsprogEngelsk
TidsskriftBioTechniques
Vol/bind68
Nummer6
Sider (fra-til)345-348
Antal sider4
ISSN0736-6205
DOI
StatusUdgivet - jun. 2020

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