ELECTRON MICROSCOPY

Sample

A17(1-16) peptide-bound singular trimer of Vaccinia virus D13

Specimen Preperation
Sample Aggregation State PARTICLE
Vitrification Instrument FEI VITROBOT MARK IV
Cryogen Name ETHANE
Sample Vitrification Details Wait time: 0sec Blot force: 0 Blot time: 5sec
3D Reconstruction
Reconstruction Method SINGLE PARTICLE
Number of Particles 1519800
Reported Resolution (Å) 2.56
Resolution Method FSC 0.143 CUT-OFF
Other Details ?
Refinement Type
Symmetry Type POINT
Map-Model Fitting and Refinement
ID 1
Refinement Space REAL
Refinement Protocol RIGID BODY FIT
Refinement Target Map-to-model correlation
Overall B Value ?
Fitting Procedure ?
Details Model building was proceeded by manually fitting the cryo-EM structure of D13 (PDB ID 7VFD) into our cryo-EM map using UCSF Chimera. The model was subsequently inspected and adjusted in Coot. Coordinate refinements were performed using the real-space refinement routine in the Phenix software suite.
Data Acquisition
Detector Type TFS FALCON 4i (4k x 4k)
Electron Dose (electrons/Å2) 42.72
Imaging Experiment
Date of Experiment ?
Temprature (Kelvin)
Microscope Model TFS KRIOS
Minimum Defocus (nm) 1200
Maximum Defocus (nm) 2200
Minimum Tilt Angle (degrees) ?
Maximum Tilt Angle (degrees) ?
Nominal CS 2.7
Imaging Mode BRIGHT FIELD
Specimen Holder Model ?
Nominal Magnification 105000
Calibrated Magnification ?
Source FIELD EMISSION GUN
Acceleration Voltage (kV) 300
Imaging Details ?
Imaging Experiment
Task Software Package Version
PARTICLE SELECTION cryoSPARC 4.5.3
CTF CORRECTION cryoSPARC 4.5.3
MODEL FITTING UCSF Chimera ?
MODEL FITTING Coot 0.9.8.92
MODEL REFINEMENT PHENIX 1.20.1_4487
INITIAL EULER ASSIGNMENT cryoSPARC 4.5.3
FINAL EULER ASSIGNMENT cryoSPARC 4.5.3
CLASSIFICATION cryoSPARC 4.5.3
RECONSTRUCTION cryoSPARC 4.5.3
Image Processing
CTF Correction Type CTF Correction Details Number of Particles Selected Particle Selection Details
NONE ?
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