An adhesive dispensing assembly includes a positioning module, a driving rod, a guiding block, a screw nut, a moving rod, and a motor. The positioning module is for positioning an adhesive cylinder. The screw nut is secured in the guiding block and engages with the driving rod by a screw thread. The moving rod has one end fastened to the guiding block and the other end for moving the adhesive cylinder. The motor drives the driving rod to rotate to move the screw nut, the guiding block, and the moving rod in one direction or another. An adhesive dispenser using the assembly is also described.