EMD-2059 Experiments and Validation

Cryo-EM structure of HBV T=4 pgRNA-filled Cp183 capsid

Single particle reconstruction
Overview of EMD-2059
Sample name: HBV T=4 pgRNA-filled Cp183 capsid
Organism: Hepatitis B virus
Related EM entries by publication: EMD-2060, EMD-2058, EMD-2057

Map parameters

Minimum density: -4.967
Maximum density: 9.949
Average density: 0.131
Standard deviation: 1.506
Recommended contour level: 2.00 (author)

Sample information

Sample name: HBV T=4 pgRNA-filled Cp183 capsid
Virus: Hepatitis B virus

Validation information

Experimental information

 

Image processing

Applied symmetry: I
Software: Auto3dem
Number of particles: 7201
CTF correction: Each particle phase-flipping

Imaging

Session 1
Electron source: FIELD EMISSION GUN
Electron dose (e/A**2): 14.0
Energy filter: Omega filter
Nominal CS (mm): 1.10
Nominal magnification: 40000.0
Defocus max (nm): 3,800.00
Defocus min (nm): 270.00
Holder model: GATAN LIQUID NITROGEN
Session 2
Electron source: FIELD EMISSION GUN
Electron dose (e/A**2): 14.0
Energy filter: Omega filter
Nominal CS (mm): 1.10
Nominal magnification: 40000.0
Defocus max (nm): 3,800.00
Defocus min (nm): 270.00
Holder model: GATAN LIQUID NITROGEN
Acquisition
Number of images: 193

Specimen

Preparation
Specimen state: particle
Buffer: 150 mM NaCl, 50 mM HEPES, 2 mM DTT
Specimen support: Quantifoil R 2/2 holey carbon 200 mesh copper grids
Vitrification
Apparatus: FEI VITROBOT MARK III
Cryogen: ETHANE
Humidity: 100
Protocol: Blot for 4 seconds before plunging