Efficient cloning of linear DNA inserts (ECOLI) into plasmids
using site-directed mutagenesis

J 2024

Efficient cloning of linear DNA inserts (ECOLI) into plasmids using site-directed mutagenesis

PACLÍKOVÁ, Petra a Jakub HARNOŠ

Základní údaje

Originální název

Efficient cloning of linear DNA inserts (ECOLI) into plasmids using site-directed mutagenesis

Autoři

PACLÍKOVÁ, Petra a Jakub HARNOŠ

Vydání

Scientific Reports, Nature Research, 2024, 2045-2322

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10605 Developmental biology

Stát vydavatele

Německo

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Impakt faktor

Impact factor: 3.900

Označené pro přenos do RIV

Ano

Kód RIV

RIV/00216224:14310/24:00137748

Organizační jednotka

Přírodovědecká fakulta

Klíčová slova anglicky

In vitro DNA assembly; DNA cloning; Mutagenesis; PCR; Plasmid-based cloning; Site-directed mutagenesis; Dishevelled

Štítky

14314010, rivok

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 25. 11. 2024 12:57, Mgr. Marie Novosadová Šípková, DiS.

Anotace

V originále

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.

Návaznosti

MUNI/J/0004/2021, interní kód MU

Název: Role of the Planar Cell Polarity and Mitochondria Crosstalk in Fuelling Cell Migration

Investor: Masarykova univerzita, Role of the Planar Cell Polarity and Mitochondria Crosstalk in Fuelling Cell Migration, MASH JUNIOR - MUNI Award In Science and Humanities JUNIOR

Citovat

PACLÍKOVÁ, Petra a Jakub HARNOŠ. Efficient cloning of linear DNA inserts (ECOLI) into plasmids using site-directed mutagenesis. Scientific Reports. Nature Research, 2024, roč. 14, č. 1, s. 1-8. ISSN 2045-2322. Dostupné z: https://doi.org/10.1038/s41598-024-72169-6.

@article{2454741,
   author = {Paclíková, Petra and Harnoš, Jakub},
   article_number = {1},
   doi = {https://doi.org/10.1038/s41598-024-72169-6},
   keywords = {In vitro DNA assembly; DNA cloning; Mutagenesis; PCR; Plasmid-based cloning; Site-directed mutagenesis; Dishevelled},
   language = {eng},
   issn = {2045-2322},
   journal = {Scientific Reports},
   title = {Efficient cloning of linear DNA inserts (ECOLI) into plasmids using site-directed mutagenesis},
   url = {https://www.nature.com/articles/s41598-024-72169-6},
   volume = {14},
   year = {2024}
}

TY  - JOUR
ID  - 2454741
AU  - Paclíková, Petra - Harnoš, Jakub
PY  - 2024
TI  - Efficient cloning of linear DNA inserts (ECOLI) into plasmids using site-directed mutagenesis
JF  - Scientific Reports
VL  - 14
IS  - 1
SP  - 1-8
EP  - 1-8
PB  - Nature Research
SN  - 20452322
KW  - In vitro DNA assembly
KW  - DNA cloning
KW  - Mutagenesis
KW  - PCR
KW  - Plasmid-based cloning
KW  - Site-directed mutagenesis
KW  - Dishevelled
UR  - https://www.nature.com/articles/s41598-024-72169-6
N2  - 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.
ER  -

PACLÍKOVÁ, Petra a Jakub HARNOŠ. Efficient cloning of linear DNA inserts (ECOLI) into plasmids using site-directed mutagenesis. \textit{Scientific Reports}. Nature Research, 2024, roč.~14, č.~1, s.~1-8. ISSN~2045-2322. Dostupné z: https://doi.org/10.1038/s41598-024-72169-6.

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