I collected two datasets of the same ligand bound structure at 2.4 and 2.1 Å resolution. The apo structure has already been solved to 2.2Å resolution in P31 (and P3121 for SeMet). After processing each dataset as both P31 and P3121 in MOSFLM, I proceeded with MR with Phaser using sequential partial search models to place either 3 copies in the ASU for P3121, or 6 copies for P31. (The previously solved apo structure had 6 copies of the same protein in the ASU). *** in summary two datasets, processed as both P31 or P3121 After running xtriage for either .mtz, I get this warning about NCS: Distance to origin : 56.728 Height relative to origin : 26.388 % Translational pseudo-symmetry is very likely present in these data. Be aware that this will change the intensity statistics and may impact subsequent analyses, and in practice may lead to higher R-factors in refinement. Regardless, I can proceed with the MR using sequential partial search models to place either 3 copies (P3121) or 6 copies (P31) with decent solutions (i.e. packing looks good upon inspection, scoring looks appropriate -> SOLU SET RFZ=2.8 TFZ=8.3 PAK=13*LLG*=-37 *LLG*=2199 as an example). However, I did get the following somewhat expected warnings: Number of known components (1) not divisible by 2: Translational *NCS* correction not applied Large non-origin Patterson peak indicates that translational *NCS* is present Upon any attempt at initial refinement (with a number of different strategies), the R-factors are hung up > 0.42 (for example in the P3121, 2.1Å case): Start R-work = 0.4966, R-free = 0.5133; Final R-work = 0.4231, R-free = 0.4477 My concern is that there is some pathology that is not being recognized (merohedral twinning) or that the NCS/pseudosymmetry is not being addressed appropriately. -- Cheers, John