ELECTRON MICROSCOPY

Sample

Type III-Bv CRISPR Complex from Dissulfurispira thermophila bound to a crRNA

Specimen Preperation
Sample Aggregation State PARTICLE
Vitrification Instrument FEI VITROBOT MARK IV
Cryogen Name ETHANE
Sample Vitrification Details ?
3D Reconstruction
Reconstruction Method SINGLE PARTICLE
Number of Particles 194795
Reported Resolution (Å) 3.00
Resolution Method FSC 0.143 CUT-OFF
Other Details cryoSPARC's non-uniform refinement with per particle CTF correction was used to generate the final reconstruction.
Refinement Type
Symmetry Type POINT
Map-Model Fitting and Refinement
ID 1
Refinement Space REAL
Refinement Protocol FLEXIBLE FIT
Refinement Target Cross-correlation coefficient
Overall B Value ?
Fitting Procedure ?
Details Initial fitting was done in ChimeraX, followed by refinement in PHENIX. COOT was used to manually edit the structure to better fit the density before a final refinement in PHENIX
Data Acquisition
Detector Type GATAN K3 (6k x 4k)
Electron Dose (electrons/Å2) 65
Imaging Experiment
Date of Experiment ?
Temprature (Kelvin)
Microscope Model FEI TALOS ARCTICA
Minimum Defocus (nm) 500
Maximum Defocus (nm) 2000
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 45000
Calibrated Magnification ?
Source FIELD EMISSION GUN
Acceleration Voltage (kV) 200
Imaging Details ?
Imaging Experiment
Task Software Package Version
PARTICLE SELECTION cryoSPARC 4.7.1
IMAGE ACQUISITION SerialEM 4.2.0
CTF CORRECTION cryoSPARC 4.7.1
MODEL FITTING UCSF ChimeraX 1.10.1
INITIAL EULER ASSIGNMENT cryoSPARC 4.7.1
FINAL EULER ASSIGNMENT cryoSPARC 4.7.1
CLASSIFICATION cryoSPARC 4.7.1
RECONSTRUCTION cryoSPARC 4.7.1
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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