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In Binding of Isaac: Rebirth there are various challenges which spawn you with equipped items.

What I'm wondering is if the levels are also fixed or vary each time you try the challenges. If I come across some really cool level run, will it be the same next time I try this challenge, or are they randomly generated like the 'normal' game?

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No, they are randomly generated like any normal run. Restarting, you can see that the map and the first room pattern always change.

Challenges typically start the player off with one or more specific items which make normal play more difficult. That's the only fixed thing on each try.

Note that new content (Achievements or Secrets), other than the Reward, cannot be unlocked. Likewise, you will not find a Donation Machine anywhere.
And if you find and collect new items they don't show in the Collection Page.

Only the Daily Challenge is pre-seeded. [Afterbirth DLC only]

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Seeds in challenges work the same way as with normal runs - you'll get a random seed each time you restart. The main difference is that most challenges lack Treasure Rooms and achievements cannot be earned.

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