ELECTRON MICROSCOPY

Sample

alpha1B and betaI/IVb microtubule

Specimen Preperation
Sample Aggregation State FILAMENT
Vitrification Instrument LEICA EM GP
Cryogen Name ETHANE
Sample Vitrification Details Double-cycled GMPCPP microtubules were diluted to 2.5uM in Brb80 buffer. 5 ul of microtubules were applied to glow-discharged cryo-EM grid and allowed to absorb for 30 sec.
3D Reconstruction
Reconstruction Method SINGLE PARTICLE
Number of Particles 1279335
Reported Resolution (Å) 3.1
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 FLEXIBLE FIT
Refinement Target Cross-correlation
Overall B Value 80
Fitting Procedure ?
Details ?
Data Acquisition
Detector Type GATAN K2 SUMMIT (4k x 4k)
Electron Dose (electrons/Å2) 56
Imaging Experiment
Date of Experiment ?
Temprature (Kelvin)
Microscope Model TFS KRIOS
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 ?
Nominal Magnification 130000
Calibrated Magnification ?
Source FIELD EMISSION GUN
Acceleration Voltage (kV) 300
Imaging Details ?
Imaging Experiment
Task Software Package Version
PARTICLE SELECTION RELION 3.0
IMAGE ACQUISITION SerialEM ?
CTF CORRECTION Gctf ?
MODEL FITTING Coot ?
MODEL FITTING ISOLDE ?
INITIAL EULER ASSIGNMENT RELION 3.0
FINAL EULER ASSIGNMENT RELION 3.0
RECONSTRUCTION RELION 3.0
MODEL REFINEMENT PHENIX 1.20.1_4487:
Image Processing
CTF Correction Type CTF Correction Details Number of Particles Selected Particle Selection Details
PHASE FLIPPING AND AMPLITUDE CORRECTION ?
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