Paclikova Petra, Harnos Jakub
Department of Experimental Biology, Faculty of Science, Masaryk University, 625 00, Brno, Czech Republic.
Sci Rep. 2024 Sep 16;14(1):21591. doi: 10.1038/s41598-024-72169-6.
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.
本研究介绍了一种用于克隆线性DNA质粒插入片段的新型经济高效技术，旨在解决与常用体外DNA组装方法相关的费用问题。具体而言，我们引入了ECOLI（线性插入片段高效克隆法），这是一种基于PCR产物的定点诱变方法。与其他已确立的体外DNA组装方法相比，我们的方法无需昂贵的合成或用于重组或限制位点的专用试剂盒。ECOLI为将长达数百个核苷酸的插入片段克隆到质粒构建体中提供了一种快速、高效且经济的替代方法，从而提高了克隆的可及性和效率。正如我们在WNT信号通路的Dishevelled基因上简要展示的那样，该方法可促进分子生物学研究。
