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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 inEscherichia coli [e.g., TOP10, DH5α, JM109, and BL21(DE3)] andBacillus subtilis for obtaining chimeric plasmids.

Simple Cloning via Direct Transformation of PCR Product (DNA Multimer) to Escherichia coli and Bacillus subtilis