Endonuclease SfiI is primarily used in molecular biology for DNA manipulation and cloning. Its ability to recognize and cleave specific DNA sequences makes it valuable for creating precise DNA fragments. Researchers employ SfiI in constructing DNA libraries, as it allows for the seamless insertion of DNA fragments into vectors without disrupting the reading frame. It is also utilized in gene mapping and genome analysis due to its unique recognition site and cleavage pattern. Additionally, SfiI is instrumental in creating recombinant DNA molecules, facilitating studies in genetic engineering and biotechnology. Its application extends to diagnostic assays, where it aids in the detection of specific DNA sequences.