Compared to the conventional viscose rayon process, Lyocell process using NMMO as a solvent provides a naturally-friendly, less toxic, relatively simple method for producing regenerated cellulose fiber with excellent properties. Existing wood pulp based Lyocell, however, shows some disadvantages such as forest destruction. So, in this study, a novel regenerated cellulose fiber, cotton linter based Lyocell, was prepared by dry jet-wet spinning with a lab scale equipment changing concentration of cellulose/NMMO solution, take-up speed, etc, which are strongly related with fiber properties and its properties were compared with wood pulp based Lyocell fiber. The tenacity and strain at break of the resultant fibers were correlated with the intrinsic fiber properties such as birefringence, crystallinity, and crystalline orientation index.