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The answer to this question seems to be common sense, but I just wanted to make sure, since I don’t know as much about the inner workings of Macs as I do PC’s. I want to know if I swap the optical drive in my iMac with a 2.5" hard drive using the ifixit adapter, can I make this new hard drive the OS disk? I would prefer using a SSD for my OS disk to improve performance. I just was hoping I could add a SSD to this optical drive position, and install the OS on it, and use the existing 1TB drive in the main position as my storage disk. I feel this would be cheaper and more efficient. If so, might there be any performance issues by using this secondary slot? Am I overthinking this? Any and all help is much appreciated!

Within the System Preferences you’ll find a control panel called Startup Disk within it you can control which disks OS to boot up from (if you have two different disks each with a copy of the OS present). Ideally you want the SSD to be using the HD’s SATA interface connection Vs your optical drives slower PATA interface if you have the OS on it.