ELECTRON MICROSCOPY

Sample

Human DHX9 bound to ATX-968 and ADP.

Specimen Preperation
Sample Aggregation State PARTICLE
Vitrification Instrument FEI VITROBOT MARK IV
Cryogen Name ETHANE
Sample Vitrification Details Sample was centrifuged at 21,000 x g for 10 minutes at 6 degrees Celsius and diluted to 0.5 mg/mL (4.34 micromolar) just before grid preparation. 3 microlitres of the sample was applied to the grid, blotted for 3 seconds with a blot force of 3, and immediately flash frozen.
3D Reconstruction
Reconstruction Method SINGLE PARTICLE
Number of Particles 985972
Reported Resolution (Å) 2.8
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 OTHER
Refinement Target ?
Overall B Value 64.98
Fitting Procedure ?
Details Initial fit was done in Chimera, followed by chain adjustments in Coot and refinement in Phenix.
Data Acquisition
Detector Type GATAN K3 BIOQUANTUM (6k x 4k)
Electron Dose (electrons/Å2) 60
Imaging Experiment
Date of Experiment ?
Temprature (Kelvin)
Microscope Model TFS KRIOS
Minimum Defocus (nm) 400
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 165000
Calibrated Magnification ?
Source FIELD EMISSION GUN
Acceleration Voltage (kV) 300
Imaging Details Preliminary grid screening was performed in a Glacios.
Imaging Experiment
Task Software Package Version
PARTICLE SELECTION cryoSPARC 4.7.0
IMAGE ACQUISITION EPU 3.4
CTF CORRECTION CTFFIND 4.1
MODEL FITTING Coot ?
MODEL REFINEMENT PHENIX ?
INITIAL EULER ASSIGNMENT RELION 5.0
FINAL EULER ASSIGNMENT RELION 5.0
CLASSIFICATION RELION 5.0
RECONSTRUCTION RELION 5.0
Image Processing
CTF Correction Type CTF Correction Details Number of Particles Selected Particle Selection Details
PHASE FLIPPING AND AMPLITUDE CORRECTION ?
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