ELECTRON MICROSCOPY

Sample

The structure of mouse cGAS in complex with a bubble DNA

Specimen Preperation
Sample Aggregation State PARTICLE
Vitrification Instrument FEI VITROBOT MARK IV
Cryogen Name ETHANE
Sample Vitrification Details Filter paper blotted with blot force 2 and blot time 2s, and plunge-frozen in liquid ethane using a Vitrobot Mark IV at 4 degree Celsius and 100% humidity.
3D Reconstruction
Reconstruction Method SINGLE PARTICLE
Number of Particles 62441
Reported Resolution (Å) 3.48
Resolution Method FSC 0.143 CUT-OFF
Other Details Particles are randomly split into two halves.
Refinement Type
Symmetry Type POINT
Map-Model Fitting and Refinement
ID 1
Refinement Space REAL
Refinement Protocol AB INITIO MODEL
Refinement Target Cross-correlation coefficient
Overall B Value 87.5
Fitting Procedure ?
Details Initial local fitting was done using UCSF ChimeraX, and then COOT was used for manual building.
Data Acquisition
Detector Type FEI FALCON IV (4k x 4k)
Electron Dose (electrons/Å2) 40
Imaging Experiment
Date of Experiment ?
Temprature (Kelvin)
Microscope Model TFS KRIOS
Minimum Defocus (nm) 1200
Maximum Defocus (nm) 1800
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 130000
Calibrated Magnification ?
Source FIELD EMISSION GUN
Acceleration Voltage (kV) 300
Imaging Details ?
Imaging Experiment
Task Software Package Version
PARTICLE SELECTION cryoSPARC v4.3.1
IMAGE ACQUISITION EPU 1.12 - 3.7
CTF CORRECTION cryoSPARC v4.3.1
MODEL FITTING UCSF ChimeraX version 1.9
INITIAL EULER ASSIGNMENT cryoSPARC v4.3.1
FINAL EULER ASSIGNMENT cryoSPARC v4.3.1
CLASSIFICATION cryoSPARC v4.3.1
RECONSTRUCTION cryoSPARC v4.3.1
MODEL REFINEMENT PHENIX 1.18.2_3874:
Image Processing
CTF Correction Type CTF Correction Details Number of Particles Selected Particle Selection Details
PHASE FLIPPING AND AMPLITUDE CORRECTION ?
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