ELECTRON MICROSCOPY

Sample

MCM2-7 DH bound to Sld3-Sld7, Cdc45 and DNA

Specimen Preperation
Sample Aggregation State PARTICLE
Vitrification Instrument FEI VITROBOT MARK IV
Cryogen Name ETHANE
Sample Vitrification Details Wait time: 30 s Blotting time: 1.5 s Blot force: +2
3D Reconstruction
Reconstruction Method SINGLE PARTICLE
Number of Particles 292336
Reported Resolution (Å) 2.9
Resolution Method FSC 0.143 CUT-OFF
Other Details For the composite MSC map, resolution was estimated with phenix.mtriage, using the unmasked model-map FSC, which gave an FSC(0.143) resolution of 2.9 Angstrom. Final focused refinement particle subsets used to generate composite map (292,336 particles): DH core (38,859 particles) Sld3 MRD1/2:Mcm4/6 (149,668 particles) Sld3-Cdc45:Mcm2/5 (52,475 particles) Sld7:Mcm6 (51,334 particles)
Refinement Type
Symmetry Type POINT
Map-Model Fitting and Refinement
ID 1
Refinement Space ?
Refinement Protocol ?
Refinement Target ?
Overall B Value ?
Fitting Procedure ?
Details ?
Data Acquisition
Detector Type FEI FALCON III (4k x 4k)
Electron Dose (electrons/Å2) 58.4
Imaging Experiment
Date of Experiment ?
Temprature (Kelvin)
Microscope Model TFS KRIOS
Minimum Defocus (nm) 1000
Maximum Defocus (nm) 3500
Minimum Tilt Angle (degrees) ?
Maximum Tilt Angle (degrees) ?
Nominal CS 2.7
Imaging Mode BRIGHT FIELD
Specimen Holder Model FEI TITAN KRIOS AUTOGRID HOLDER
Nominal Magnification 81000
Calibrated Magnification ?
Source FIELD EMISSION GUN
Acceleration Voltage (kV) 300
Imaging Details ?
Imaging Experiment
Task Software Package Version
PARTICLE SELECTION Topaz ?
IMAGE ACQUISITION EPU ?
CTF CORRECTION CTFFIND 4.1.14
INITIAL EULER ASSIGNMENT RELION 4.0
FINAL EULER ASSIGNMENT cryoSPARC 4.6.2
CLASSIFICATION cryoSPARC 4.6.2
RECONSTRUCTION cryoSPARC 4.6.2
RECONSTRUCTION RELION 4.0
RECONSTRUCTION PHENIX 1.21.2-5419
MODEL REFINEMENT PHENIX 1.21.2_5419
Image Processing
CTF Correction Type CTF Correction Details Number of Particles Selected Particle Selection Details
PHASE FLIPPING AND AMPLITUDE CORRECTION ?
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