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Title:Characterization of the extremophilic, archaeal virus STIV2
Authors:L.J.Happonen,P.Redder,X.Peng,L.J.Reigstad,D.Prangishvili,S.J.Butcher
Sample:STIV2 virus
Aggregation state:Icosahedral (20 angstroms resolution)
Red flagLatest update:2011-09-09
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Sample
Sample name: STIV2 virus
Components:
ID Type Name Exp. MW (MDa) Oligomeric details Recombinant expression Synthetic Organism GO identifier InterPro identifier Virus identifier Details
1virusSulfolobus turreted icosahedral virus 2Sulfolobus turreted icosahedral virus 2
Experiment
Sample preparation:
pHSample conc.DetailsStainingSample support details
3.5 mg/mL50 mM sodium citrate, pH 3.5Vitrified. Grids were blotted for roughly one second before being plunged into liquid ethane.Quantifoil-grids
Vitrification:
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
ETHANE% KHOMEMADE PLUNGERA small vial of ethane is placed inside a larger liquid nitrogen reservoir. The grid holding 3 microliters of the sample is held in place at the bottom of a plunger by the means of fine tweezers. When the liquid ethane is ready, a piece of filter paper is then pressed against the sample to blot off excess buffer, sufficient to leave a thin layer on the grid. The filter paper is removed, and the plunger is allowed to drop into the liquid ethane. Once the grid enters the liquid ethane, the sample is rapidly frozen, and the grid is transferred under liquid nitrogen to a storage box immersed in liquid nitrogen for later use in the microscope. msVitrification instrument: Guillotine
Imaging:
MicroscopeVoltageIllumination modeImaging modeCsDefocus min.Defocus max.Nominal mag.Calibrated mag.Electron sourceDetectorDetector distanceAstigmatism
FEI TECNAI F20200 kVFLOOD BEAMBRIGHT FIELD2.0 mm500 nm5200 nm6800066400FIELD EMISSION GUNGatan Ultrascan 4000 CCD camera mm

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
Side entry liquid nitrogen-cooled side entry holderGATAN LIQUID NITROGEN°° eV K K K mrad18 e/Å2Low dose conditions
Processing
Software:PFT, POR, EM3DR2, P3DR
CTF correction:Each micrograph
Resolution by author:20 Å
Resolution method:FSC at 0.5 cut-off
Unit cell:
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
3584.42 μm/pixellink
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails
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