Endonuclease NsiI is widely utilized in molecular biology for DNA manipulation and analysis. It is particularly valuable in restriction enzyme mapping, where it cleaves DNA at specific recognition sites, aiding in the identification of genetic sequences. This enzyme is also employed in cloning experiments to create compatible ends for ligation, facilitating the insertion of DNA fragments into vectors. Additionally, NsiI is used in diagnostic applications, such as genotyping and mutation detection, by enabling precise cutting of DNA at targeted locations. Its specificity and efficiency make it a crucial tool in genetic research and biotechnology.