Efficient cloning of linear DNA inserts (ECOLI) into plasmids using site-directed mutagenesis
Jazyk angličtina Země Velká Británie, Anglie Médium electronic
Typ dokumentu časopisecké články
Grantová podpora
MUNI/J/0004/2021
Grant Agency of Masaryk University
PubMed
39284917
PubMed Central
PMC11405386
DOI
10.1038/s41598-024-72169-6
PII: 10.1038/s41598-024-72169-6
Knihovny.cz E-zdroje
- Klíčová slova
- Dishevelled, DNA cloning, In vitro DNA assembly, Mutagenesis, PCR, Plasmid-based cloning, Site-directed mutagenesis,
- MeSH
- DNA genetika MeSH
- klonování DNA * metody MeSH
- mutageneze cílená * metody MeSH
- plazmidy * genetika MeSH
- polymerázová řetězová reakce metody MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- DNA MeSH
This study introduces a novel cost-effective technique for cloning of linear DNA plasmid inserts, aiming to address the associated expenses linked with popular in vitro DNA assembly methods. Specifically, we introduce ECOLI (Efficient Cloning Of Linear Inserts), a method utilizing a PCR product-based site-directed mutagenesis. In comparison to other established in vitro DNA assembly methods, our approach is without the need for costly synthesis or specialized kits for recombination or restriction sites. ECOLI offers a fast, efficient, and economical alternative for cloning inserts up to several hundred nucleotides into plasmid constructs, thus enhancing cloning accessibility and efficiency. This method can enhance molecular biology research, as we briefly demonstrated on the Dishevelled gene from the WNT signaling pathway.
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