EMD-2246 Experiments and Validation

Cryo-electron microscopy of phirsl1 jumbo phage

Single particle reconstruction
Overview of EMD-2246
Sample name: phirsl1 baseplate
Organism: Ralstonia phage RSL1
Related EM entries by publication: EMD-2244, EMD-2245, EMD-2247, EMD-2243

Map parameters

Minimum density: -0.06
Maximum density: 0.07
Average density: 0
Standard deviation: 0
Recommended contour level: 0.01 (author)

Sample information

Sample name: phirsl1 baseplate
Protein: phirsl1

Validation information

Experimental information

 

Image processing

Applied symmetry: C6
Software: SPIDER
Number of particles: 1228
Reconstruction protocol: projection matching
CTF correction: each particle

Imaging

Session
Microscope model: FEI POLARA 300
Electron source: FIELD EMISSION GUN
Electron dose (e/A**2): 20.0
Nominal CS (mm): 2.3
Nominal magnification: 31000.0
Defocus max (nm): 5,295
Defocus min (nm): 1,355
Holder model: GATAN LIQUID NITROGEN
Acquisition
Number of images: 52
Scanner model: ZEISS SCAI
Sampling interval (μm): 7.0

Specimen

Preparation
Specimen state: particle
Buffer: 50 mM Tris-HCl [pH 7.5], 100 mM NaCl, 10 mM MgSO4
Staining procedure: Four microliters of the bacteriophage sample (~0.1 mg/ml) were loaded between the mica-carbon interface. The sample was stained using 2% ammonium molybdate pH 7.5 and air-dried.
Vitrification
Apparatus: FEI VITROBOT MARK IV
Cryogen: ETHANE
Humidity: 100