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

Rasmus Hejlesen; Ernst Martin Füchtbauer

doi:10.2144/btn-2019-0104

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

Rasmus Hejlesen, Ernst Martin Füchtbauer

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avis › Tidsskriftartikel › Forskning › peer review

4 Citationer (Scopus)

Abstract

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.

Originalsprog	Engelsk
Tidsskrift	BioTechniques
Vol/bind	68
Nummer	6
Sider (fra-til)	345-348
Antal sider	4
ISSN	0736-6205
DOI	https://doi.org/10.2144/btn-2019-0104
Status	Udgivet - jun. 2020

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10.2144/btn-2019-0104Licens: CC BY-NC-ND

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abstract = "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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AU - Hejlesen, Rasmus

AU - Füchtbauer, Ernst Martin

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AB - 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.

KW - donor template generation

KW - gene editing

KW - multiple site-directed mutagenesis

KW - prolonged overlap extension

KW - seamless cloning

KW - seamless plasmid assembly

UR - http://www.scopus.com/inward/record.url?scp=85086524917&partnerID=8YFLogxK

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DO - 10.2144/btn-2019-0104

M3 - Journal article

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VL - 68

SP - 345

EP - 348

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

Abstract

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