ELECTRON MICROSCOPY

Sample

NHE9 homodimer

Specimen Preperation
Sample Aggregation State PARTICLE
Vitrification Instrument FEI VITROBOT MARK IV
Cryogen Name ETHANE
Sample Vitrification Details Sample vol 3ul; blot force -5; Blot time 7s; Wait time 30sec
3D Reconstruction
Reconstruction Method SINGLE PARTICLE
Number of Particles 212666
Reported Resolution (Å) 2.83
Resolution Method FSC 0.143 CUT-OFF
Other Details Final refinement using non-uniform refinement
Refinement Type
Symmetry Type POINT
Map-Model Fitting and Refinement
ID 1
Refinement Space REAL
Refinement Protocol FLEXIBLE FIT
Refinement Target ?
Overall B Value ?
Fitting Procedure ?
Details Initial model fitted and manually rebuilt/refined in COOT and final refinement in ISOLDE and PHENIX
Data Acquisition
Detector Type TFS FALCON 4i (4k x 4k)
Electron Dose (electrons/Å2) 50
Imaging Experiment
Date of Experiment ?
Temprature (Kelvin)
Microscope Model TFS KRIOS
Minimum Defocus (nm) 1000
Maximum Defocus (nm) 2400
Minimum Tilt Angle (degrees) ?
Maximum Tilt Angle (degrees) ?
Nominal CS ?
Imaging Mode BRIGHT FIELD
Specimen Holder Model FEI TITAN KRIOS AUTOGRID HOLDER
Nominal Magnification 165000
Calibrated Magnification ?
Source FIELD EMISSION GUN
Acceleration Voltage (kV) 300
Imaging Details ?
Imaging Experiment
Task Software Package Version
PARTICLE SELECTION Topaz 0.2.4
PARTICLE SELECTION cryoSPARC 3.3.1
IMAGE ACQUISITION EPU ?
CTF CORRECTION cryoSPARC 3.3.1
MODEL FITTING Coot 0.9.8.93
MODEL FITTING ISOLDE 1.3
INITIAL EULER ASSIGNMENT cryoSPARC 3.3.1
FINAL EULER ASSIGNMENT cryoSPARC 3.3.1
CLASSIFICATION cryoSPARC 3.3.1
RECONSTRUCTION cryoSPARC 3.3.1
MODEL REFINEMENT PHENIX 1.21.1_5286
Image Processing
CTF Correction Type CTF Correction Details Number of Particles Selected Particle Selection Details
PHASE FLIPPING AND AMPLITUDE CORRECTION ?
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