At setup time, yes.
https://support.yubico.com/hc/en-us/articles/360016614880-Ca...
Look at the "few exceptions" section
https://support.yubico.com/hc/en-us/articles/360016614880-Ca...
Look at the "few exceptions" section
So one strategy for maintaining multiple authenticators or yubikeys would be to save all the secrets away and call them up when it's time to load up the alternate factor.
What I do is register both keys at the same time, one after the other so the QR code doesn't leave my screen. Can't do it this way if the one key is in cold storage, of course.