Restriction Endonucleases: Molecular Cloning and Beyond | NEB

The sequence-specific DNA cleavage activity of restriction endonucleases (REases), combined with other enzymatic activities that amplify and ligate nucleic acids, have enabled modern molecular biology. After more than half a century of research and development, the applications of REases have evolved from the cloning of exogenous DNA and genome mapping to more sophisticated applications, such as the identification and mapping of epigenetic modifications and the high-throughput assembly of combinatorial libraries. Furthermore, the discovery and engineering of nicking endonucleases (NEases) has opened the door to techniques such as isothermal amplification of DNA among others. In this review, we will examine the major breakthroughs of REase research, applications of REases and NEases in various areas of biological research and novel technologies for assembling large DNA molecules.