Sample information
Sample name: T. thermophilus 70S ribosome
Ribosome:
70S ribosome
Validation information
Experimental information
Image processing
Applied
symmetry: C1
Software: CTFFIND3, RELION
Number of
particles: 35404
CTF
correction: each image
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
Fitting
Fitting 1
Initial
atomic model:
2WH1
Fitting
software: Chimera
Fitting
protocol: rigid body
Refinement
space: REAL
Details:
Protocol: Rigid body. The domains were separately fitted by manual docking using program Chimera
Fitting 2
Initial
atomic model:
2WH2
Fitting
software: Chimera
Fitting
protocol: rigid body
Refinement
space: REAL
Details:
Protocol: Rigid body. The domains were separately fitted by manual docking using program Chimera
Imaging
Session
Microscope
model: FEI POLARA 300
Electron
source: FIELD EMISSION GUN
Electron
dose (e/A**2): 16.0
Nominal
CS (mm):
2.00
Nominal
magnification: 59000.0
Defocus
max (nm):
3,597.00
Defocus
min (nm):
1,178.00
Holder
model: GATAN LIQUID NITROGEN
Acquisition
Number
of images: 360
Sampling
interval (μm):
14.0
Details:
Every image is the average of sixteen frames recorded by the direct electron detector
Specimen
Preparation
Specimen
state: particle
Specimen
concentration (mg/mL):
0.20
Buffer:
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
Specimen
support: Quantifoil grids (2/2) with 3 nm thin carbon on top
Vitrification
Apparatus:
FEI VITROBOT MARK II
Cryogen:
ETHANE
Humidity:
100
Protocol:
Blot 2.5 seconds before plunging