EMD-8585

Single-particle
4.05 Å
EMD-8585 Deposition: 02/02/2017
Map released: 05/04/2017
Last modified: 16/08/2017
Overview 3D View Sample Experiment Validation Volume Browser Additional data Links
Overview 3D View Sample Experiment Validation Volume Browser Additional data Links

EMD-8585

CryoEM structure of crosslinked E.coli RNA polymerase elongation complex

EMD-8585

Single-particle
4.05 Å
EMD-8585 Deposition: 02/02/2017
Map released: 05/04/2017
Last modified: 16/08/2017
Overview 3D View Sample Experiment Validation Volume Browser Additional data Links
Method: Single-particle
Aggregation State: Particle
Specimen preparation [1]
Concentration: 3 mg/mL
Details: This sample was monodisperse
Buffer
pH: 8.0
Buffer components [4]:
Name Formula Concentration ChEBI
Potassium chloride KCl 150.0 mM
Tris Tris 20.0 mM
Magnesium chloride MgCl2 5.0 mM
Dithiothreitol DTT 5.0 mM
Grid
Model: C-flat CF-1.2/1.3-3Au
Material: GOLD
Pretreatment
Type: GLOW DISCHARGE
Atmosphere:
Support Film [1]
Material Topology Thickness
CARBON HOLEY -
Vitrification
Cryogen name: ETHANE
Chamber humidity: 100%
Chamber temperature: 295 K
Instrument: FEI VITROBOT MARK IV
Microscopy [1]
Microscope: FEI TITAN KRIOS
Illumination mode: FLOOD BEAM
Imaging mode: BRIGHT FIELD
Electron source: FIELD EMISSION GUN
Acceleration voltage: 300 kV
Nominal CS: 2.7 mm
Nominal defocus: 0.8 µm - 2.4 µm
Calibrated defocus: 0.8 µm - 2.5 µm
Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER
Image Recording [1]
Detector model: GATAN K2 SUMMIT (4k x 4k)
Detector mode: SUPER-RESOLUTION
Dimensions: 3710 pixel x 3838 pixel
Frames per image: 3-50
Number of grids: 1
Number of real images: 2000
Average exposure time: 15.0 s
Average electron dose per image: 85.2 e/Å2
Image processing [1]
Final reconstruction
Resolution: 4.05 Å ( BY AUTHOR)
Resolution method: FSC 0.143 CUT-OFF
Number of classed used: 1
Number of images used: 83800
Applied Symmetry
Point group: C1
Software [1]
Name Version Details
RELION 2.0 -
Startup model [1]
Type: OTHER
Details:E.coli core enzyme structure were extracted from PDB 4LLG, and converted to map by using EMAN2 command e2pdb2mrc.py
Initial angle assignment
Type: OTHER
Software [1]
Name Version Details
RELION 1.4 -
Final angle assignment
Type: OTHER
Software [1]
Name Version Details
RELION 1.4 -
Particle selection [1]
Selected Ref. model Method Software Details
275600 - - - -
Final 3D classification
Number of classes: 6
Avg. number of members per classes: 45900.0
Software [1]
Name Version Details
RELION 2.0 -
CTF correction
Details:CTF parameters were estimated by CTFFIND4
Software [1]
Name Version Details
CTFFIND4 4 -
Map
Format: CCP4
Data type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Annotation details: CryoEM structure of crosslinked E.coli RNA polymerase elongation complex
Geometry
X Y Z
Dimensions 256 256 256
Origin 0 0 0
Spacing 256 256 256
Voxel size 1.3 Å 1.3 Å 1.3 Å
Contour list
Primary Level Source
True 0.015 EMDB