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Title:Zernike Phase Contrast electron cryo-tomography of cyanophage Syn5 assembly intermediates
Authors:Dai W, Fu C, Raytcheva D, Flanagan J, Khant HA, Liu X, Rochat RH, Haase-Pettingell C, Piret J, Ludtke SJ, Nagayama K, Schmid MF, King JA, Chiu W
Sample:Cyanophage Syn5 assembly intermediate inside cyanobacteria Synechococcus WH8109 cells
Method:Subtomogram averaging (52 angstroms resolution)
Red flagLatest update:2013-11-06
Sample name: Cyanophage Syn5 assembly intermediate inside cyanobacteria Synechococcus WH8109 cells
ID Type Name Exp. MW (MDa) Theo. MW (MDa) Oligomeric details Recombinant expression Synthetic Organism UniProt identifier GO identifier InterPro identifier Virus identifier Details
1virusSynechococcus phage Syn5Synechococcus phage Syn5DNA-containing capsid with tail and horn - assembly intermediate
Specimen state: Particle
Specimen preparation:
pHSpecimen conc.DetailsStainingSpecimen support details
8.0 mg/mLartificial sea water200 mesh Quantifoil holey grids, glow discharged
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
ETHANE95%85 KFEI VITROBOT MARK IIIBlot for 3 seconds before plunging ms
MicroscopeVoltageIllumination modeImaging modeCsDefocus min.Defocus max.Nominal mag.Calibrated mag.Electron sourceDetectorDetector distanceAstigmatism
JEOL 2200FS200 kVFLOOD BEAMBRIGHT FIELD4.1 mm0 nm1000 nm2500025000FIELD EMISSION GUNGATAN ULTRASCAN 4000 (4k x 4k)1200 mmObjective lens astigmatism was corrected at 100,000 times magnification

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
GATAN LIQUID NITROGEN-60°60°in-column GIF energy filter15-20 eV103 K93 K105 K mrad1 e/Å201-JUN-2011
Protocol:back projection
Number of subtomogram averaged:96
Number of class averages:1
Imposed symmetry:C1
Resolution by author:52 Å
Resolution method:FSC 0.143
Processing details:The subtomograms were extracted from infected cells.
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
42 μm/pixel16
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails