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Appl Environ Microbiol. 2012 Mar;78(5):1593-5. doi: 10.1128/AEM.07105-11. Epub 2011 Dec 22.

Simple cloning via direct transformation of PCR product (DNA Multimer) to Escherichia coli and Bacillus subtilis.

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  • 1Biological Systems Engineering Department, Virginia Polytechnic Institute and State University, Blacksburg, VA, USA.


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.

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