简单克隆通过直接转化 PCR 产物（DNA 多聚体）到大肠杆菌和枯草芽孢杆菌。
Simple cloning via direct transformation of PCR product (DNA Multimer) to Escherichia coli and Bacillus subtilis.
机构信息
Biological Systems Engineering Department, Virginia Polytechnic Institute and State University, Blacksburg, VA, USA.
出版信息
Appl Environ Microbiol. 2012 Mar;78(5):1593-5. doi: 10.1128/AEM.07105-11. Epub 2011 Dec 22.
Abstract
We developed a general restriction enzyme-free and ligase-free method for subcloning up to three DNA fragments into any location of a plasmid. The DNA multimer generated by prolonged overlap extension PCR was directly transformed in Escherichia coli [e.g., TOP10, DH5α, JM109, and BL21(DE3)] and Bacillus subtilis for obtaining chimeric plasmids.
摘要
我们开发了一种通用的无需限制酶和连接酶的方法，可将多达三个 DNA 片段亚克隆到质粒的任何位置。通过长片段重叠延伸 PCR 产生的 DNA 多聚体可直接转化到大肠杆菌[如 TOP10、DH5α、JM109 和 BL21(DE3)]和枯草芽孢杆菌中，以获得嵌合质粒。
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