amplification of whole large plasmids via transposon-mediated insertion.

DNA amplification is a fundamental technique in molecular biology. The replication cycle reaction is a new method for amplification of large circular DNA having sequences, which is a replication initiation site of the chromosome. We here developed a replication cycle reaction-based method useful for amplification of various circular DNAs lacking , even in the absence of any sequence information, via transposon-mediated insertion to the circular DNA template. A 15-kb non- plasmid was amplified from a very small amount of starting DNA (50 fg, 1 fM). The method was also applicable to GC-rich plasmid (69%) or large F-plasmid (230 kb). This method thus provides a powerful tool to amplify various environmental circular DNAs.