ELECTRON MICROSCOPY

Sample

Ribosome fraction isolated from wild type P. aeruginosa cells during exponential phase growth

Specimen Preperation
Sample Aggregation State PARTICLE
Vitrification Instrument FEI VITROBOT MARK IV
Cryogen Name ETHANE
Sample Vitrification Details Blot time: 5 seconds Blot force: 3
3D Reconstruction
Reconstruction Method SINGLE PARTICLE
Number of Particles 642285
Reported Resolution (Å) 2.25
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 ?
Overall B Value ?
Fitting Procedure ?
Details Initially, each chain of the 7UNW 50S subunit was rigid body fit into the experimental map. Occasionally Alpha fold 2 generated models of individual chains were used instead. Models were then refined in phenix.
Data Acquisition
Detector Type GATAN K3 (6k x 4k)
Electron Dose (electrons/Å2) 54.3
Imaging Experiment
Date of Experiment ?
Temprature (Kelvin)
Microscope Model FEI TALOS ARCTICA
Minimum Defocus (nm) 500
Maximum Defocus (nm) 2500
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 36000
Calibrated Magnification ?
Source FIELD EMISSION GUN
Acceleration Voltage (kV) 200
Imaging Details ?
Imaging Experiment
Task Software Package Version
PARTICLE SELECTION cryoSPARC v4.7.1
IMAGE ACQUISITION SerialEM 4.1
CTF CORRECTION cryoSPARC v4.7.1
MODEL FITTING Coot 0.9.5
MODEL REFINEMENT PHENIX 1.20.1_4487
INITIAL EULER ASSIGNMENT cryoSPARC v4.7.1
FINAL EULER ASSIGNMENT cryoSPARC v4.7.1
CLASSIFICATION cryoSPARC v4.7.1
RECONSTRUCTION cryoSPARC v4.7.1
Image Processing
CTF Correction Type CTF Correction Details Number of Particles Selected Particle Selection Details
PHASE FLIPPING AND AMPLITUDE CORRECTION ?
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