Letter reSeArCH
741.854 Particles1.506.073 Particles picked
(WARP)2 Rounds of 2D classication with
2-fold binned particles (1,96 Å pixel size)192.527 Particles
25,95 %47.559 Particles
6,41 %66.265 Particles
8,93 %66.942 Particles
9,02 %368.561 Particles
49,68 %368.561 Particles
4,52 Å105.011 Particles
27,8 %155.444 Particles
43,8 %108.106 Particles
28,4 %108.106 Particles
4,52 Å3D renement19.851 Particles
18,64 %65.743 Particles
59,76 %22.512 Particles
21,16 %3D classication3D classication3D classication19.851 Particles
4,15 ÅNo image alignment; T = 819.851 Particles
4,15 ÅRe-extraction with
full pixel size of 0,96 Å3D renement3D renement3D renement Bayesian polishingCTF and beam
tilt renementExtended Data Fig. 7 | Cryo-EM single-particle analysis pipeline.
Data-processing strategy for cryo-EM of the SidJ–CaM complex. Particle-
picking on 2,423 micrographs (using WARP) resulted in the identification
of 1,500,000 particles. The particle coordinates were imported into
Relion and particles were extracted with a twofold binning factor. After
2D classification and an initial 3D classification, a 3D class with clear
secondary structure features and 370,000 particles was identified. The
particles of this class were re-extracted with full pixel size and 3D-refined,
resulting in a 4.52-Å model. Two additional rounds of 3D classification
and 3D refinement improved the resolution of the model to 4.15 Å. Final
particle polishing and CTF refinement of the remaining particles did not
result in a nominal improvement of the resolution.