The invention discloses a tool set with a magnetic head separation supporting frame and for disassembly and transplantation of a 2.5-cun mechanical hard disk magnetic head assembly and a use method of the tool set and relates to disassembly and transplantation of the 2.5-cun mechanical hard disk magnetic head assembly. The tool set with the magnetic head separation supporting frame and for disassembly and transplantation of the 2.5-cun mechanical hard disk magnetic head assembly solves the problems that due to the fact that an existing tool is not provided with a device and an instrument which are used for holding the magnetic head assembly, a magnetic head support deforms due to scraping and collision or adhesion of a magnetic head occurs and solves the problem that due to the fact that a use method of the existing tool is complicated and difficult, and the work efficiency is low. One end of a handle of the magnetic head separation supporting frame is fixed to the bottom of a separation assembly of the magnetic head support. A magnetic head assembly holding platform comprises a platform base, a magnetic head assembly fixing rod, a magnetic head assembly tray and two front-end drive circuit brackets, wherein the magnetic separation supporting frame is arranged in a groove of the platform base. The magnetic head separation supporting frame is used for disassembly of the magnetic head assembly, and the disassembled magnetic head assembly is placed on the magnetic head assembly holding platform. The tool set is suitable for disassembly and transplantation of the 2.5-cun mechanical hard disk magnetic head assembly.