Hi All, I want to apologize in advance for such a basic question. I'm working with a tetrameric protein and am attempting to build a model. The resolution estimate of the map determined by cryoEM is 3.8A. Some parts of the map are great while some other parts not so much. To assist with modelling the tetramer, I used the local refinement of one protomer to build a protomer model (one chain). I then used that protomer model as a template to build the full tetrameric model, and I make sure that each protomer is exactly the same (using ChimeraX or coot) by copying one chain over to the next and making certain they are all exact copies of one another. When doing real space refinement in phenix 2.0-5936 of the tetramer model, I end up with good statistics and what I think is a complete model. RCSB/comprehensive cryoEM validation also seem to complement my modelling. However upon closer inspection I noticed that many rotamers across the model are different, for example Chains A and B will have a Gln in one position, while chains C and D will have rotamers in totally different positions. These rotameric seem to happen in substandard regions of map quality. My understanding is that all the protomers in a C4 channel should look exactly the same. To attempt to rectify this problem I've made sure that NQH_flips is off (rotameric changes still happen with many different residues), as well as tried using reference restraints of my protomer, or NCS constraints of one protomer (these options results in a degradation of model quality). I'm missing or misunderstanding something here (or maybe a combo of both), and I was wondering if there was any advice from the community on how to proceed. Kind Regards, Seb