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Title:Cryo-electron microscopy of phirsl1 jumbo phage
Authors:Effantin G, Hamasaki R, Kawasaki T, Bacia M, Moriscot C, Weissenhorn W, Yamada T, Schoehn G
Sample:phirsl1 baseplate
Method:Single particle reconstruction (25 angstroms resolution)
Sample
Sample name: phirsl1 baseplate
Components:
ID Type Name Exp. MW (MDa) Theo. MW (MDa) Oligomeric details Recombinant expression Synthetic Organism UniProt identifier GO identifier InterPro identifier Virus identifier Details
1proteinphirsl1falseRalstonia phage RSL1
Experiment
Specimen state: Particle
Specimen preparation:
pHSpecimen conc.DetailsStainingSpecimen support details
7.5 mg/mL50 mM Tris-HCl [pH 7.5], 100 mM NaCl, 10 mM MgSO4Four 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:
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
ETHANE100% KFEI VITROBOT MARK IV ms
Imaging:
MicroscopeVoltageIllumination modeImaging modeCsDefocus min.Defocus max.Nominal mag.Calibrated mag.Electron sourceDetectorDetector distanceAstigmatism
FEI POLARA 300300 kVSPOT SCANBRIGHT FIELD2.3 mm1355 nm5295 nm31000FIELD EMISSION GUNKODAK SO-163 FILM mmObjective lens astigmatism was corrected at 59000

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
GATAN LIQUID NITROGEN°° eV K K K mrad20 e/Å225-FEB-2011
Processing
Protocol:projection matching
Software:SPIDER
CTF correction:each particle
Number of particles:1229
Imposed symmetry:C6
Resolution by author:25 Å
Resolution method:FSC 0.5
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
527 μm/pixel8ZEISS SCAI
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails