The Gateway^® cloning system sets a new trend in molecular biology by addressing the difficulties of adaptability, efficiency and compatibility of the traditional cloning approaches. Based on the well-characterized site-specific recombination system of phage lambda, the Gateway^® technology allows the cloning, combining and transferring of DNA segments between different expression platforms in a high-throughput manner while maintaining orientation and the reading frame of the fragment or fragments of interest. In this article, the key-aspects and potential applications of this system are reviewed. Benefits and weaknesses of this and other cloning technologies are discussed.