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Title: Ribosome structures to near-atomic resolution from thirty thousand cryo-EM particles Authors: Bai XC, Fernandez IS, McMullan G, Scheres SHW Sample: S.cereviseae 80S ribosome Method: Single particle reconstruction (4.6 angstroms resolution)   Summary  Experimental details   Visualization   Map information   Downloads Sample Sample name: S.cereviseae 80S ribosome Theoretical molecular weight of the sample: 4.2 Components: ID Type Name Exp. MW (MDa) Oligomeric details Recombinant expression Synthetic Organism UniProt identifier GO identifier InterPro identifier Virus identifier Details 1 ribosome-eukaryote 80S ribosome 4.2 false Saccharomyces cerevisiae Experiment Specimen state: Particle Specimen preparation: pH Specimen conc. Details Staining Specimen support details 7.45 0.3 mg/mL 3mM Hepes-KOH, 6.6 mM Tris-acetate pH 7.2, 3 mM NH4Cl, 6.6 mM NH4-acetate, 48 mM K-acetate, 4 mM Mg-acetate, 2.4 mM DTT Quantifoil grids (2/2) with 3 nm thin carbon on top Vitrification: Cryogen name Humidity Temp. Instr. Method Time resolved Details ETHANE 100% 90 K FEI VITROBOT MARK II Blot 2.5 seconds before plunging ms Imaging: Microscope Voltage Illumination mode Imaging mode Cs Defocus min. Defocus max. Nominal mag. Calibrated mag. Electron source Detector Detector distance Astigmatism FEI POLARA 300 300 kV FLOOD BEAM BRIGHT FIELD 2 mm 1191 nm 3440 nm 59000 79096 FIELD EMISSION GUN FEI Falcon (backthinned) mm Objective lens astigmatism was corrected at 59,000 times magnification Specimen holder Holder model Tilt min. Tilt max. Energy filter Energy window Temp. Temp. min. Temp. max. Beam tilt Electron dose Other details Date GATAN LIQUID NITROGEN ° ° eV 85 K 80 K 90 K mrad 16 e/Å2 15-JUL-2012 Processing Software: CTFFIND3, RELION CTF correction: each image Number of particles: 22638 Resolution by author: 4.6 Å Resolution method: gold-standard FSC at 0.143 cut-off Processing details: Use a newly developed statistical movie processing approach to compensate for beam-induced movement. Use a newly developed statistical movie processing to compensate for beam-induced movement. Scanned images: Num. images Sampling size OD range Quant. bit number Other details Scanner 200 14 μm/pixel Every image is the average of sixteen frames recorded by the direct electron detector Fitting: PDB Protocol Target crit. Software B value Fitting space PDB chain Details 3U5B  rigid body Chimera REAL Protocol: Rigid body. The domains were separately fitted by manual docking using program Chimera 3U5C  rigid body Chimera REAL Protocol: Rigid body. The domains were separately fitted by manual docking using program Chimera 3U5D  rigid body Chimera REAL Protocol: Rigid body. The domains were separately fitted by manual docking using program Chimera 3U5E  rigid body Chimera REAL Protocol: Rigid body. The domains were separately fitted by manual docking using program Chimera