Single-molecule PCR using water-in-oil emulsion.


Polymerase chain reaction (PCR) using a single molecule of DNA is very useful for analysis, detection and cloning of the desired DNA fragment. We developed a simple PCR method utilizing a water-in-oil (W/O) emulsion that included numerous droplets of reaction mixture in bulk oil phase. These droplets, which were stable even at high temperatures, functioned as micro-reactors. This allows the effective concentration of template DNA to be increased, even for low concentrations of template DNA. The present method consists of a two-step thermal cycle. The first step was carried out using the W/O emulsion. During this step, the template DNA was amplified in the limited volume of the droplets in the W/O emulsion. The W/O emulsion was broken and the second PCR step was carried out. This method can be easily applied to amplify a single DNA molecule.

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@article{Nakano2003SinglemoleculePU, title={Single-molecule PCR using water-in-oil emulsion.}, author={Michihiko Nakano and Jun Komatsu and Shun-ichi Matsuura and Kazunori Takashima and Shinji Katsura and Akira Mizuno}, journal={Journal of biotechnology}, year={2003}, volume={102 2}, pages={117-24} }